Allergen dosage form

ABSTRACT

The invention provides allergen containing pharmaceutical products and in particular fast-dispersing solid allergen dosage forms. In particular, fast-dispersing, non-compressed solid dosage forms suitable for oromucosal administration comprising a matrix and at least one allergen are provided. Suitable matrices are gelatine, starch and mannitol. Methods for the dosage forms are also provided.

This application is a divisional of U.S. patent application Ser. No.10/723,308, filed Nov. 26, 2003, which claims the benefit of U.S.Provisional Application Ser. No. 60/429,086 filed Nov. 26, 2002, DanishPatent Application PA 200201852, filed Nov. 26, 2002, and Danish PatentApplication PA 200300279 filed Feb. 24, 2003. Each of these priorityapplications is incorporated herein by reference and in their entirety.

FIELD OF THE INVENTION

This invention relates to an allergen containing pharmaceutical productand in particular to fast-dispersing solid allergen dosage forms and amethod for preparing such dosage forms.

BACKGROUND OF THE INVENTION

Allergy is a major health problem in countries where Western lifestyleis adapted. Furthermore, the prevalence of allergic disease isincreasing in these countries. Although allergy in general may not beconsidered a life-threatening disease, asthma annually causes asignificant number of deaths. An exceptional prevalence in about 30% ofteenagers conveys a substantial loss in quality of life, working daysand money, and warrants a classification among major health problems inthe Western world.

Allergy is a complex disease. Many factors contribute to thesensitisation event. Among these is the susceptibility of the individualdefined by an as yet insufficiently understood interplay between severalgenes. Another important factor is allergen exposure above certainthresholds. Several environmental factors may be important in thesensitisation process including pollution, childhood infections,parasite infections, intestinal microorganisms, etc. Once an individualis sensitised and the allergic immune response established, the presenceof only minute amounts of allergen is efficiently translated intosymptoms.

The natural course of allergic disease is usually accompanied byaggravation at two levels. First, there is a progression of symptoms anddisease severity. For example, there is a progression from hay fever toasthma. Secondly, dissemination in offending allergens most often occursresulting in allergic multi-reactivity. Chronic inflammation leads to ageneral weakening of the mucosal defense mechanisms resulting inunspecific irritation and eventually destruction of the mucosal tissue.Infants may become sensitised primarily to foods, i.e. milk, resultingin eczema or gastrointestinal disorders; however, most often theyoutgrow these symptoms spontaneously. These infants are at risk ofdeveloping inhalation allergy later in their lives.

The most important allergen sources are found among the most prevalentparticles of a certain size in the air we breathe. These sources areremarkably universal and include grass pollens and house dust mitefaecal particles, which together are responsible for approximately 50%of all allergies. Of global importance are also animal dander, i.e. catand dog dander, other pollens, such as mugwort pollens, and micro-fungi,such as Alternaria. On a regional basis other pollens may dominate, suchas birch pollen in Northern and Central Europe, ragweed in the Easternand Central United States, and Japanese cedar pollen in Japan. Insects,i.e. bee and wasp venoms, and foods each account for approximately 2% ofall allergies.

Allergy, i.e. type I hyper-sensitivity, is caused by an inappropriateimmunological reaction to foreign non-pathogenic substances. Importantclinical manifestations of allergy include asthma, hay fever, eczema,and gastro intestinal disorders. The allergic reaction is prompt andpeaks within 20 minutes upon contact with the offending allergen.Furthermore, the allergic reaction is specific in the sense that aparticular individual is sensitised to particular allergen(s), whereasthe individual does not necessarily show an allergic reaction to othersubstances known to cause allergic disease. The allergic phenotype ischaracterized by a pronounced inflammation of the mucosa of the targetorgan and by the presence of allergen specific antibody of the IgE classin the circulation and on the surfaced of mast-cells and basophils.

An allergic attack is initiated by the reaction of the foreign allergenwith allergen specific IgE antibodies, when the antibodies are bound tohigh affinity IgE specific receptors on the surface of mast-cells andbasophils. The mast-cells and basophils contain preformed mediators,i.e. histamine, tryptase, and other substances, which are released uponcross-linking of two or more receptor-bound IgE antibodies. IgEantibodies are cross-linked by the simultaneous binding of one allergenmolecule. The cross-linking of receptor bound IgE on the surface ofmast-cells also leads to release of signalling molecules responsible forthe attraction of eosinophils, allergen specific T-cells, and othertypes of cells to the site of the allergic response. These cells ininterplay with allergen, IgE and effector cells, lead to a renewed flashof symptoms occurring 12-24 hours after allergen encounter (late phasereaction).

Allergy disease management comprises diagnosis and treatment includingprophylactic treatments. Diagnosis of allergy is concerned with thedemonstration of allergen specific IgE and identification of theallergen source. In many cases a careful anamnesis may be sufficient forthe diagnosis of allergy and for the identification of the offendingallergen source material. Most often, however, the diagnosis issupported by objective measures, such as skin prick test, blood test, orprovocation test.

The therapeutic options fall in three major categories. The firstopportunity is allergen avoidance or reduction of the exposure. Whereasallergen avoidance is obvious e.g. in the case of food allergens, it maybe difficult or expensive, as for house dust mite allergens, or it maybe impossible, as for pollen allergens. The second and most widely usedtherapeutic option is the prescription of classical symptomatic drugslike anti-histamines and steroids. Symptomatic drugs are safe andefficient; however, they do not alter the natural cause of the disease,and they do not control the disease dissemination. The third therapeuticalternative is specific allergy vaccination that in most cases reducesor alleviates the allergic symptoms caused by the allergen in question.

Conventional specific allergy vaccination is a causal treatment forallergic disease. It interferes with basic immunological mechanismsresulting in persistent improvement of the patients' immune status.Thus, the protective effect of specific allergy vaccination extendsbeyond the treatment period in contrast to symptomatic drug treatment.Some patients receiving the treatment are cured, and in addition, mostpatients experience a relief in disease severity and symptomsexperienced, or at least an arrest in disease aggravation. Thus,specific allergy vaccination has preventive effects reducing the risk ofhay fever developing into asthma, and reducing the risk of developingnew sensitivities.

The immunological mechanism underlying successful allergy vaccination isnot known in detail. A specific immune response, such as the productionof antibodies against a particular pathogen, is known as an adaptiveimmune response. This response can be distinguished from the innateimmune response, which is an unspecific reaction towards pathogens. Anallergy vaccine is bound to address the adaptive immune response, whichincludes cells and molecules with antigen specificity, such as T-cellsand the antibody producing B-cells. B-cells cannot mature into antibodyproducing cells without help from T-cells of the correspondingspecificity. T-cells that participate in the stimulation of allergicimmune responses are primarily of the Th2 type. Establishment of a newbalance between Th1 and Th2 cells has been proposed to be beneficial andcentral to the immunological mechanism of specific allergy vaccination.Whether this is brought about by a reduction in Th2 cells, a shift fromTh2 to Th1 cells, or an up-regulation of Th1 cells is controversial.Recently, regulatory T-cells have been proposed to be important for themechanism of allergy vaccination. According to this model regulatoryT-cells, i.e. Th3 or Tr1 cells, down-regulate both Th1 and Th2 cells ofthe corresponding antigen specificity. In spite of these ambiguities itis generally believed that an active vaccine must have the capacity tostimulate allergen specific T-cells, preferably TH1 cells.

Primarily for two reasons specific allergy vaccination in spite of itsvirtues, is not in widespread use. One reason is the inconveniencesassociated with the traditional vaccination programme that comprisesrepeated vaccinations, such as injections over several months. The otherreason is, more importantly, the risk of allergic side reactions.Ordinary vaccinations against infectious agents are efficientlyperformed using a single or a few high dose immunizations. Thisstrategy, however, cannot be used for allergy vaccination since apathological immune response is already ongoing.

Conventional specific allergy vaccination is therefore carried out usingmultiple subcutaneous immunizations applied over an extended timeperiod. The course is divided in two phases, the up dosing and themaintenance phase. In the up dosing phase increasing doses are applied,typically over a 16-week period, starting with minute doses. When therecommended maintenance dose is reached, this dose is applied for themaintenance phase, typically with injections every six weeks. Followingeach injection the patient must remain under medical attendance for 30minutes due to the risk of anaphylactic side reactions, which inprinciple although extremely rare could be life-threatening. Inaddition, the clinic should be equipped to support emergency treatment.There is no doubt that a vaccine based on a different route ofadministration would eliminate or reduce the risk for allergic sidereactions inherent in the current subcutaneous based vaccine as well aswould facilitate a more widespread use, possibly even enabling selfvaccination at home.

Attempts to improve vaccines for specific allergy vaccination have beenperformed for over 30 years and include multifarious approaches. Severalapproaches have addressed the allergen itself through modification ofthe IgE reactivity. Others have addressed this route of administration.

The immune system is accessible through the oral cavity and oromucosal,e.g. sublingual administration, of allergens is a known route ofadministration.

Conventionally, allergy vaccine using the oromucosal route consists ofthe periodic dosing of a solution of the allergen at intervals spacedapart by at least one day. In comparison, the therapeutic (accumulated)maintenance doses given exceeded the maintenance of the comparablesubcutaneous dose by a factor of 5-500. Obvious drawbacks of this dosageform and route of administration are the problems associated withaccurate and uniform self administration of the correct dose by thepatient (several drops may have to be given, uniformity of theindividual drops, application site accuracy, etc.). Additionally, thereis a need to refrigerate the drug and include preservatives in theformulation.

Netien et al. (“Galenica 16—Médicaments homéopathiques” ed. 2, 1986,pages 77-99) discloses a liquid solution impregnated onto a solidparticulate (granules) or conventional compressed tablets of lactose,saccharose or a mixtures of these for sublingual administration ofmedicaments such as allergens.

DD-A.0 107 208 discloses a process for preparing a conventionalcompressed tablet containing an allergen. Upon administration the tabletis dissolved by the saliva and the allergen is then absorbed through themucosa of the oral cavity. The formulation contains a water insolubleexcipient, namely talcum as well as paraffin and fatty acids which isnot desirable because it will leave an unpleasant remnant in the mouthof the patient. Moreover, the friction produced during the tablettingprocess may be detrimental to the physical stability of the allergens

EP 278 877 discloses a pharmaceutical composition for sublingual use,where a solid support is coated with a solution of an allergen whenspraying the solution onto solid support globules. The resultingformulation is alleged to disintegrate rapidly, but not instantaneously.However, there is no disclosure of how to achieve the objective.Moreover, the formulation contains reducing sugars in the form oflactose, which are prone to react with allergens.

In order to ensure that as much as possible of an administered dose of acertain allergen is presented to the mucosa of the oral cavity andadditionally that the contact time of the disintegrated product with themucosa is maximised, it is very important that the dosage formdisintegrates instantaneously upon contact with the saliva of the oralcavity. Fast dispersing solid dosage forms, which readily release theactive ingredient in the oral cavity are known in the art.

U.S. Pat. No. 4,371,516 discloses pharmaceutical dosage forms containingactive ingredients, which disintegrate rapidly in water. Thepharmaceutical dosage forms comprise an open matrix network of carriermaterial, which disintegrate within 10 seconds.

A freeze-dried fish gelatine based carrier as disclosed in WO 00/61117is designed to release the active ingredient instantaneously uponcontact with saliva when administered in the oral cavity.

A freeze-dried modified starch carrier as disclosed in WO 00/44351 isdesigned to release the active ingredient instantaneously upon contactwith saliva when administered in the oral cavity.

WO 99/21579 discloses a fast-dispersing dosage form comprising a vaccineand an adjuvant for oral use.

WO 02/13858 discloses fast dissolving pharmaceutical compositioncontaining vaccines in the form of a fast dissolving “cake” for oraluse. The object of WO 02/13858 appears to be to provide viral orbacterial vaccines that will stay intact in the gastrointestinal tract.This is achieved by protecting the antigen against the acidic content ofthe stomach by incorporating antacids such as calcium carbonate into thecake.

WO 00/51568 discloses a fast-disintegrating compressed low friabilitytablet that is designed to dissolve in the mouth in contact with salivain less than 30 seconds forming an easy-to-swallow suspension.

It is alleged in U.S. Pat. No. 4,371,516 that the formulation is usefulfor oral vaccines. In the case of WO 00/61117, WO 00/44351, WO 99/21579and WO 02/13858, it is also alleged that the inventions are directed tonon-infections immuno-modulated conditions such as systemic allergicconditions e.g. hayfever. However, there is no disclosure in any ofthese applications of technical information or examples of how afast-dispersing allergen vaccine solid dosage form can be manufactured.For example, there are no indications of an appropriate dosage of acertain allergen in any of the disclosed formulation. It is veryimportant to administer a correct dosage of an allergen to a patient,because an excess dose may induce anaphylactic shock in the patient.Furthermore, no recognition of or indications of appropriate measures inrelation to stability or friability of such formulations are given.

SUMMARY OF THE INVENTION

The present invention concerns a pharmaceutical product suitable fororomucosal administration of an allergen comprising at least oneallergen and a matrix in form of a fast-dispersing non-compressed soliddosage form characterized in that the allergen containing dosage form isstable, sufficiently robust and does not release hazardous amounts ofallergen residues upon handling by the patient.

Further, a fast-dispersing non-compressed solid dosage form suitable fororomucosal administration comprising a matrix forming agent and anallergen wherein the allergen is stable and the dosage form has a lowfriability with respect to allergen release, the dosage form dissolvesquickly and does not require an adjuvant.

In particular the invention concerns a pharmaceutical product suitablefor administration of allergen comprising

-   -   a fast-dispersing, non-compressed solid dosage form suitable for        oromucosal administration including:        -   a) a matrix formed from at least one matrix-forming agent,            and        -   b) an effective dose of an allergen for desensitizing an            individual to said allergen,    -   wherein        -   (c) the loss of the allergen content in said dosage form is            less than 50% of the initial allergen content after being            held for 3 months at 25° C. and 60% relative humidity, and        -   (d) the loss of allergen from said solid dosage form is less            than about 0.5 μg allergen extract or less than about 0.05            μg major allergen when subjected to a friability test.

In one preferred embodiment of the invention, the solid dosage formcomprises fish gelatine and mannitol as matrix-forming agents.

In another preferred embodiment of the invention, the solid dosage formcomprises starch and mannitol as matrix forming agents.

The present invention also provides methods of producing these soliddosage forms and methods of treating allergy by administration of thesesolid dosage forms.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based on a number of surprising findings, noneof which could with a reasonable expectation of success be predicted apriori. First, it is based on the finding that it is possible to use afast-dispersing, non-compressed dosage form for administration of anallergen, and that it is possible to obtain an effective allergytreatment by the use of such a formulation. In particular, it has beenshown that it is indeed possible to obtain delivery of a sufficientamount of allergen to the immune system of the patient via theoromucosal route using a fast-dispersing non-compressed solid dosageform without eliciting an undesirable level of side effects. Moreover,the present invention has provided the relevant dose levels forobtaining a therapeutic effect without unacceptable side effects.

Secondly, the invention is based on the finding that an effectivetreatment can be obtained by the use of a fast-dispersing,non-compressed dosage form without the use of an adjuvant.

Thirdly, the invention is based on the finding that it is possible toformulate an allergen as a fast-dispersing, non-compressed dosage formwhile meeting the special demands required by an allergen with respectto stability and low friability. In particular, it has been shown thatit is possible to balance the oppositely directed requirements that thedosage form should be fast-dispersing on the one side and both stableand have low-friability on the other side so as to obtain a dosage form,which to a sufficient extent is fast-dispersing, stable and has lowfriablility.

Allergen proteins are susceptible to degradation that is influenced by anumber of factors of the environments they are contained in. It issought that it is pertinent for allergy treatment and in particular toallergy vaccination that the allergen is delivered intact to the immunesystem in therapeutically relevant doses. Thus, the allergen must remainstable during production, storage and use. The present work has shownthat it is in fact possible to formulate allergen proteins in afast-dispersing, non-compressed dosage form which is stable in respectto allergen doses and allergen activity. Moreover, it has surprisinglybeen found that these formulations are indeed stable at roomtemperature. This finding has significant importance for the handlingprocedures of the final product. Cold storage at the manufacturingplant, during transport or during storage at the pharmacy is oftenassociated with high cost, because the cooling facilities have to beclosely monitored and it is very expensive to invest in reliable coolingfacilities. Moreover, with respect to compliance of the patient, it isalso preferable that the dosage form can be stored at room temperature.

Thus, present work has shown that it is in fact possible to formulateallergen proteins in a fast-dispersing, non-compressed dosage form beingtherapeutically effective without the need for an adjuvant. Further suchallergen containing solid dosage forms are furthermore stable atadvantageous conditions.

When allergen proteins are formulated in a fast-dispersing,non-compressed dosage form, it is further advantageous that theresulting dosage form does not substantially release the allergens tothe surrounding environment or to a person handling the dosage form upondirect contact. A priori non-compressed fast-dispersing solid dosageforms are characterized by a low mechanical strength compared tocompressed tablets due to the inherent nature of the non-compressedmatrix, which is fragile and brittle, almost wafer-like. During e.g.packing, storage, transport and handling of the dosage form by thepatient, residual particles containing the allergen may be released tothe environment and patient. This is especially detrimental when theactive ingredient is an allergen, because the allergen can elicit anallergic reaction in a disposed person or induce an allergic reaction,such sensitisation or allergic response being dose dependent. Maximumallowable levels for environmental contamination in the form of e.g.allergen in dust have been proposed, depending on the allergen inquestion, to be as low as 2 micrograms major allergen per gram housedust. (Allergy. Principles and practice (1993, 4. ed.), Mosby-Year book,Vol. I page 520).

Non-compressed fast-dispersing solid dosage forms, which aremanufactured by removal of a liquid from a solidified system comprisingmatrix forming agents, active ingredient and other optional agents,preferably are manufactured in situ. The in situ manufacturing processgenerally involves removal of solvent from a solidified system of theactive ingredient and the matrix forming excipients within the finalcontainer such as a blister pack. The in situ technique usedcommercially does not allow for conventional coating of the dosage form.Application of a coating of the dosage form would in most cases affectdispersing of the solid dosage form, thus jeopardizing the instantaneousrelease properties of the dosage form.

Therefore, despite the opposite nature of these requirements, thepresent work has shown that it is in fact possible to formulateeffective dosages of allergen proteins in a fast-dispersing,non-compressed dosage form, while at the same time obtaininglow-friability and maintaining the fast-dispersing property.

All of the above findings are supported by experimental laboratory workor pre-clinical experiments using test animals, where applicable, orclinical trials, where applicable.

The term “fast-dispersing dosage form” refers to dosage forms whichdisintegrate in less than about 90 seconds, preferably in less thanabout 60 seconds, preferably in less than about 30 seconds, morepreferably in less than about 20, even more preferably in less thanabout 10 seconds in the oral cavity, even more preferred in less thanabout 5 seconds, and most preferably in less than about 2 seconds afterbeing received in the oral cavity. The solid dosage form of theinvention may be in the form of tablets, capsules, lozenges or caplets.

The term “non-compressed” refers to a solid dosage form, which ismanufactured by removal of a liquid from a solidified system comprisingmatrix forming agents, active ingredient and other suitable ingredientsresulting in an allergen comprised solid matrix.

The term “solid dosage form” refers to a unit dosage form that is not aliquid, or a powder when it is administered in the oral cavity, thus“solid dosage forms” refers to e.g. tablets containing a unit dose ofthe active ingredient.

The term tablets, solid dosage and vaccine form are used interchangeablyherein.

The term “matrix forming agent” refers to any pharmaceuticallyacceptable water-soluble or water-dispersible excipient that will serveas a carrier for the active ingredient in the solid dosage form.

The term “excipient” refers to any ingredient that may be added to theformulation besides the active ingredient.

The term “loss of the allergen content in said dosage form” refers tofor example degradation or inactivation of the allergen in the dosageform during for example storage, transportation and use. The loss may bedetermined as either the loss in biological acticivity/potency or as theloss in the actual content of the allergen. Preferably, the loss of theallergen content is measured as the loss of at least one major allergen.The loss may for example be measured in an ELISA method as described inObispo et al. (Allergy, 1997, 52, pg. 806-813 using allergen specificreagents).

The term “loss of the allergen content from said dosage form” refers tofor example release of the allergen in the dosage form during, forexample storage, transportation and use. The loss may be determined aseither the loss in biological acticivity/potency or as the loss in theactual content of the allergen. Preferably, the loss of the allergencontent is measured as the loss of at least one major allergen. The lossmay for example be measured in an ELISA method as described in Obispo etal. (Allergy, 1997, 52, pg. 806-813 using allergen specific reagents).

The term “stable” refers to dosage forms where the loss in allergencontent is less than 50% of the initial content after being stored for 3months at 25° C. and 60% relative humidity in the final container eithermeasured as the loss in biological acticivity/potency or as the loss incontent of at least one major allergen. The loss may for example bemeasured in an ELISA method as described above.

The term “low friability” refers to the amount of allergen containingmaterial that is lost from the dosage form when it is subjected to anexternal force. The solid dosage form has a sufficient friability androbustness to be transported, stored and handled if the allergencontaining material lost contains less than 0.5 μg allergen extract or0.05 μg major allergen per solid dosage form. For the purpose of thepresent invention, the friability may be measured by a method accordingto the present invention.

“Tensile strength σ” is calculated according to the following equation:σ=3Wa×9.8 Nmm⁻²/2d ² b

where w=Peak load to fracture (kgF)

a=distance between supports

d=thickness of the fast-dispersing solid dosage form (mm)

b=diameter of the fast-dispersing solid dosage form (mm)

“Peak load to fracture” means the peak force required to fracture a unitin a three point bend test using an appropriate instrument (e.g. CT5,Engineering Systems, 1 Loach Court, Radford Bridge Road, Nottingham NG81NA, UK).

The term “oromucosal administration” refers to a route of administrationwhere the dosage form is placed under the tongue or anywhere else in theoral cavity to allow the active ingredient to come in contact with themucosa of the oral cavity or the pharynx of the patient in order toobtain a local or systemic effect of the active ingredient. An exampleof an oromucosal administration route is sublingual administration.

The term “sublingual administration” refers to a route ofadministration, where a dosage form is placed underneath the tongue inorder to obtain a local or systemic effect of the active ingredient.

The term “allergen” refers to any naturally occurring protein ormixtures of proteins that have been reported to induce allergic, i.e.IgE mediated reactions upon their repeated exposure to an individual.Examples of naturally occurring allergens include pollen allergens(tree, weed, herb and grass pollen allergens), mite allergens (from e.g.house dust mites and storage mites), insect allergens (inhalant, saliva-and venom origin allergens), animal allergens from e.g. saliva, hair anddander from e.g. dog, cat, horse, rat, mouse, etc., fungi allergens andfood allergens. The allergen may be used in the form of an allergenextract, a purified allergen, a modified allergen or a recombinantallergen or a recombinant mutant allergen, any allergen fragment above30 amino acids or any combination thereof.

The expression “allergen extract” as used therein refers to an extractobtained by extraction of a biological allergen source material asgenerally described in “Allergenic extracts”, H. Ipsen et al, chapter 20in Allergy, principle and practise (Ed. S. Manning) 1993, Mosby-YearBook, St. Louis. Such extract may be obtained by aqueous extraction ofwater soluble material followed by purification steps like filtration toobtain the solution i.e. the extract. The extract may then be subjectedto further purification and/or processing like freeze-drying removingsubstantially all the water. Generally, an allergen extract comprises amixture of proteins and other molecules. Allergen proteins are oftenclassified as a major allergen, an intermediate allergen, a minorallergen or no classification. An allergen extract generally comprisesboth major and minor allergens. Major allergens will generallyconstitute approximately 5-15% of an average allergen extract, moreoften about 10%. Classification of an allergen is based on an assessmentof the clinical importance of the particular allergen and is givenbelow. Examples of important major allergen found in an extract includegrass group 1 and 5 and 6 allergens (e.g. Phl p 1, 5, and 6), dust mitegroup 1 and 2 allergens (e.g. Der p 1, Der p 2), tree pollen allergen 1(Bet v 1), cedar pollen allergen 1 and 2 (e.g. Cry j 1, Cry j 2),ragweed pollen 1 and 2 (Amb a 1, Amb a 2), cat allergen 1 (i.e. Fel d1).The average allergic person will be sensitised to and react to one ormore major allergens and further may also be sensitised and react tominor allergens.

Amounts of allergen extract referred to herein refers to the dry mattercontent of such allergen extracts.

Preferably the water content of the dry matter does not exceed 10%, morepreferably 5% by weight.

The expression “biological allergen source material” as used thereinrefers to any biological material comprising one or more allergens.Examples of such materials are acarids PMB (Pure Mite Body) or WMC(Whole Mite Culture), defatted or non-defatted pollens from e.g.grasses, herbs, weeds and trees, animal hair and dander, pelt, fungimycelia and spores, insect bodies, venom or saliva and foods.

Biological allergen source materials may comprise contaminatingmaterials, such as foreign pollen and plant and flower debris for anallergen pollen source material.

The degree of contamination should be minimised. Preferably, the contentof contaminants should not exceed 10% (W/W) of the biological sourcematerial.

Normally an allergen extract contains at least 10% protein of the drymatter content of the allergen extract as determined in a standardprotein assay such as BCA or Lowry and the remainder consists of other“non-protein material,” which may be components such as lipids,carbohydrates, or bound water which originate from the biologicalallergen source.

An allergen extract may be formulated and stored in form of afreeze-dried material obtainable by freeze-drying a liquid allergenextract at a pressure of below 800 micro bar and for a period of up till100 hours removing the water.

In the field of allergy extracts, there is no international acceptedstandardisation method. A number of different units of extract strengthi.e. bio-potency exist. The methods employed and the units used normallymeasure the allergen content and biological activity. Examples hereofare SQ-Units (Standardised Quality units), BAU (Biological AllergenUnits), BU (biological units), UM (Units of Mass), IU (InternationalUnits) and IR (Index of Reactivity). Hence, if extracts of origins otherthan those disclosed herein are used, they need to be standardisedagainst extract disclosed herein in order to determine their potency inSQ units or any of the above mentioned units. The subject matter isdealt with in “Allergenic extracts”, H. Ipsen et al, chapter 20 inAllergy, principle and practise (Ed. S. Manning) 1993, Mosby-Year Book,St. Louis and Løwenstein H. (1980) Arb Paul Ehrlich Inst 75:122.

The bio-potency, i.e. the in vivo allergenic activity, of a givenextract depends on a number of factors, the most important being thecontent of major allergens in the extract, which varies with thecomposition of the biological source material.

The amount of allergen extract in grams to be used for obtaining adesired bio-potency varies with the type of extract in question, and fora given type of extract the amount of allergen extract varies from onebatch to another with the actual bio-potency of the extract.

For a given batch of extract, the amount of allergen extract in grams tobe used for obtaining a desired bio-potency may be determined using thefollowing procedure:

-   -   (a) The bio-potency of various amounts of a reference extract is        determined using one or more immunological in vivo tests to        establish a relationship between bio-potency and amount of        reference extract. Examples of the said immunological in vivo        tests are Skin Prick Test (SPT), Conjunctival Provocation Test        (CPT), Bronchial Challenge with Allergen (BCA) and various        clinical trials in which one or more allergy symptoms is        monitored, see for example e.g. Haugaard et al., J Allergy Clin        Immunol, Vol. 91, No. 3, pp 709-722, March 1993.    -   (b) On the basis of the established relationship between        bio-potency and reference extract, the bio-potency of one or        more relevant doses for use in the dosage forms of the invention        is selected with due consideration to a balance of the factors        of i) the effect of treating or alleviating symptoms of        allergy, ii) side effects recorded in the immunological in vivo        tests, and iii) the variability of i) and ii) from one        individual to another. The balancing is done to obtain a maximal        adequate therapeutic effect without experiencing an unacceptable        level of side effect. The way of balancing the factors are well        known to those skilled in the art

The bio-potency of the one or more relevant doses found may be expressedin any biopotency unit available, such as SQ units, BAU, IR units, IU,cf. above.

-   -   (c) From the reference extract one or more bio-potency reference        standard extracts is prepared and, if used, the bio-potency unit        values of the reference standard extracts are calculated on the        basis of the bio-potency unit value allocated to the one or more        relevant doses, e.g. such a standard for BAU can be obtained        from FDA as illustrated below.    -   (d) For the reference standard extracts of each extract type, a        number of parameters for evaluating the bio-potency of extracts        are selected. Examples of such evaluation parameters are total        allergenic activity, the amount of defined major allergens and        overall molecular composition of the extract. The total        allergenic activity may be measured using an in vitro        competitive immunoassay, such as ELISA and MagicLite®        luminescence immunoassay (LIA), using a standardised antibody        mixture raised against the extract obtained using standard        methods, e.g. antibodies raised in mouse or rabbit, or a pool of        allergic patients sera. The content of major allergens may e.g.        be quantified by rocket immuno-electrophoresis (RIE) and        compared to the reference standards. The overall molecular        composition may be examined using e.g. crossed        immunoelectrophoresis (CIE) and sodium dodecyl sulphate        polyacrylamide gel electrophoresis (SDS-PAGE).    -   (e) For a given batch of extract of unknown bio-potency (test        extract), the amount of extract to be used for obtaining a        desired bio-potency level (effective dose for use in the solid        dosage form according to the present invention) may be        determined as follows: For each evaluation parameter selected,        the test extract is compared with the reference standard        extracts using the relevant measurement methods as described        above, and on the basis of the measurement results the amount of        extract having the desired bio-potency is calculated.

SQ-Unit: The SQ-Unit is determined in accordance with ALK-Abelló A/S's“SQ biopotency”-standardisation method, where 100,000 SQ units equal thestandard subcutaneous maintenance dose. Normally 1 mg of extractcontains between 100,000 and 1,000,000 SQ-Units, depending on theallergen source from which they originate and the manufacturing processused. The precise allergen amount can be determined by means ofimmunoassay i.e. total major allergen content and total allergenactivity.

BAU (Biological Allergen Units) is biological potency units asdetermined according to the requirements of the FDA for allergen productdescribed in “Quantitative determination of relative potency ofallergenic extracts” (“Methods of the allergen products testingLaboratory” “ELISA competition assay”. Page 15, #49N-0012, FDA, October1993). A dose of 100,000 SQ-Units containing grass extract equals acontent of 2600-4700 BAU according to the method above. Likewise, otherextracts can be assessed according to the method above.

The term “effective dose of an allergen for desensitization” shall meana dose which when taken once or repeatedly in a monodose or inincremental doses results in, for example, an adaptive immune responseand thus serves as means to desensitise allergic patients. Preferably,the term shall mean the amount of allergen in each dosage form necessaryto induce an adaptive immune response after repeated administration ofsaid solid dosage forms in accordance with a treatment regimen (over aperiod ranging from a few applications to at least one daily applicationover several months). Preferably desensitization includes thealleviation of allergic symptoms upon administration of the dose.Clinical allergy symptoms include rhinitis, conjunctivitis, asthma,urticaria, eczema, which includes reactions in the skin, eyes, nose,upper and lower airways with common symptoms such as redness and itchingof eyes and nose, itching and runny nose, coaching, weezing, shortnessof breathe, itching, and swelling of tissue.

“Uniformity of content” as used herein refers to the variation of thedoses unit from the stated dose.

“Water content” as used herein refers to the content of residual waterin a solid dosage unit determined quantitatively using the Karl Fischertitration principle. This method is based on the principle that a givenamount of I₂ leads to a transformation of an equivalent amount of water(European Pharmacopoeia (EP) 3^(rd) edition, 2.5.12).

As used herein “Water activity a_(w)” is the effective water in asample. Water activity measurements are carried out using methods knownto the person skilled in the art, for example chilled mirror dew pointtechnology, relative humidity with sensors that change electricalresistance or capacitance or using a lithium chloride electrode:

a_(w) can be calculated according to the following equation:a _(w) =p/ps=ERH (%)/100

where

-   -   p=partial pressure of water vapor at the surface of the product    -   ps=saturation pressure, or the partial pressure of water vapor        above pure water at the product temperature.    -   ERH=equilibrium relative humidity.

The term “about” or “approximately” means within an acceptable range forthe particular value as determined by one of ordinary skill in the art,which will depend in part on how the value is measured or determined,e.g., the limitations of the measurement system. For example, “about”can mean a range of up to 20%, preferably up to 10%, more preferably upto 5%, and more preferably still up to 1% of a given value.

It has now surprisingly been found that solid dosage form according tothe invention provides an oromucosal pharmaceutical allergen product,which provides effective allergen doses giving an allergen specificimmune response in a dose-response manner and having acceptable sideeffects.

It has further been found that it is indeed possible to manufacture anon-compressed fast-dispersing solid dosage form with low friabilitycontaining allergens, which is sufficiently robust and does not releasehazardous amounts of residue upon handling by the patient.

Moreover, it has surprisingly been found that these formulations arestable at room temperature.

Allergens are in a varying degree particularly susceptible todegradation in an aqueous environment, such as in an aqueous allergensolution or in a product with high water content and/or high wateractivity.

The European Pharmacopoeia monograph for Allergen Products and “Note forguidance on allergen products”, CPMP (London 13, March 1996) state thatthe moisture levels should not exceed 5% for freeze-dried products (i.e.allergen extracts in vials), and that these should be stored frozen(−20° C.). Cold storage (2-8° C.) is also a requirement for liquid-basedsublingual formulations, which further have a limited shelf life. It hassurprisingly been found that allergens including labile allergens arestable at room temperature conditions. Even the dosage forms accordingto the invention having water content above the prescribed maximum levelof 5% are stable at room temperature. Without being bound to theory,this may be explained by the fact that the excipients of thefast-dispersing solid dosage form bind the remaining water in the dosageform and reduces the water activity of the allergen solid dosage form.Hence, by reducing the water activity of the formulation, it is possibleto obtain a stable formulation with no degradation of the allergen, eventhough the water content is higher than the maximum level of 5% that isprescribed for allergen extracts in vials.

Water activity is one important factor contributing to the shelf life ofa product. It is well known that the water activity of a product affectsgrowth of bacteria as well as the stability, the potency and consistencyof pharmaceuticals. Also protein stability is influenced significantlyby water activity due to the relatively fragile nature of proteins. Mostproteins must maintain conformation to remain active. Maintaining lowwater activity levels helps to prevent or entice conformational changes,which subsequently is important to ensure that a protein in the form ofan allergen is stable. Also hydrolytic degradation of proteins, whethercaused by enzymes or not, is affected by the water activity.

Further it is believed that the water content will affect the mechanicalstrength of the solid dosage forms. In general, high values willincrease the risk of the solid dosage form becoming more deliquescent,whereas lower value will affect the robustness of the solid dosage form,e.g. the solid dosage becoming more friable and brittle.

Water activity measurements are carried out by using methods known tothe person skilled in the art for example chilled mirror dew pointtechnology, relative humidity with sensors that change electricalresistance or capacitance or using a lithium chloride electrode.

The water activity of a solid dosage form preferable does not exceed0.70 and preferably is between 0.1-0.7, more preferably is between0.2-0.6, more preferably is between 0.3-0.5, and most preferably isbetween 0.4-0.5.

The water content of a solid dosage form determined according to themethod described in Example 1 preferably does not exceed 25% andpreferably is between 0.1%-20%, more preferably is between 0.5-15%, morepreferably is between 2-8%, more preferably is between 4-7%, mostpreferably between 4.5-6% water.

According to one embodiment of the invention an allergen pharmaceuticalproduct is provided in a fast-dispersing solid dosage form, whichrapidly dissolves in the oral cavity on contact with saliva, hencebringing the allergen in close contact with the immunological relevanttissue of the mucosa and allowing the allergen to address these.Examples of naturally occurring allergens include pollen allergens(tree, herb, weed, and grass pollen allergens), insect allergens(inhalant, saliva and venom allergens, e.g. mite allergens, cockroachand midges allergens, hymenoptera venom allergens), animal hair anddander allergens (from e.g. dog, cat, horse, rat, mouse, etc.), and foodallergens. Important pollen allergens from trees, grasses and herbs aresuch originating from the taxonomic orders of Fagales, Oleales, Pinalesand Platanaceae including for example birch (Betula), alder (Alnus),hazel (Corylus), hornbeam (Carpinus) and olive (Olea), cedar(Cryptomeria and Juniperus), Plane tree (Platanus), the order of Poalesincluding for example grasses of the genera Lolium, Phleum, Poa,Cynodon, Dactylis, Holcus, Phalaris, Secale, and Sorghum, the orders ofAsterales and Urticales including for example herbs of the generaAmbrosia, Artemisia, and Parietaria. Other important inhalationallergens are those from house dust mites of the genus Dermatophagoidesand Euroglyphus, storage mite e.g. Lepidoglyphys, Glycyphagus andTyrophagus, those from cockroaches, midges and fleas e.g. Blatella,Periplaneta, Chironomus and Ctenocepphalides, and those from mammalssuch as cat (genus Felis), dog (genus Canis), cow (genus Bos) and horse(genus Equus), venom allergens including such originating from stingingor biting insects such as those from the taxonomic order of Hymenopteraincluding bees (superfamily Apidae), wasps (superfamily Vespidea), andants (superfamily Formicoidae). Important inhalation allergens fromfungi are, for example, those originating from the genera Alternaria andCladosporium.

In a more preferred embodiment of the invention the allergen is Bet v 1,Aln g 1, Cor a 1 and Car b 1, Que a 1, Cry j 1, Cry j 2, Cup a 1, Cup s1, Jun a 1, Jun a 2, jun a 3, Ole e 1, Lig v 1, Pla l 1, Pla a 2, Amb a1, Amb a 2, Amb t 5, Art v 1, Art v 2 Par j 1, Par j 2, Par j 3, Sal k1, Ave e 1, Cyn d 1, Cyn d 7, Dac g 1, Fes p 1, Hol l 1, Lol p 1 and 5,Pha a 1, Pas n 1, Phl p 1, Phl p 5, Phl p 6, Poa p 1, Poa p 5, Sec c 1,Sec c 5, Sor h 1, Der f 1, Der f 2, Der p 1, Der p 2, Der p 7, Der m 1,Eur m 2, Gly d 1, Lep d 2, Blo t 1, Tyr p 2, Bla g 1, Bla g 2, Per a 1,Fel d 1, Can f 1, Can f 2, Bos d 2, Equ c 1, Equ c 2, Equ c 3, Mus m 1,Rat n 1, Apis m 1, Api m 2, Ves v 1, Ves v 2, Ves v 5, Dol m 1, Dol m 2,Dol m 5, Pol a 1, Pol a 2, Pol a 5, Sol i 1, Sol i 2, Sol i 3 and Sol i4, Alt a 1, Cla h 1, Asp f 1, Bos d 4, Mal d 1, Gly m 1, Gly m 2, Gly m3, Ara h 1, Ara h 2, Ara h 3, Ara h 4, Ara h 5 or shufflant hybrids fromMolecular Breeding (Maxygen, Inc.) of any of these.

In the most preferred embodiment of the invention the allergen is grasspollen allergen or a dust mite allergen or a ragweed allergen or a cedarpollen or a cat allergen or birch allergen.

In yet another embodiment of the invention the fast-dispersing soliddosage form comprises at least two different types of allergens eitheroriginating from the same allergenic source or originating fromdifferent allergenic sources. For example the fast-dispersing soliddosage form comprises grass group 1, grass group 2/3, grass group 5 andgrass group 6 allergens or mite group 1 and group 2 allergens fromdifferent mite and grass species respectively, weed antigens like shortand giant ragweed allergens, different fungi allergens like alternariaand cladosporium, tree allergens like birch, hazel, hornbeam, oak andalder allergens, food allergens like peanut, soybean and milk allergens.

The allergen incorporated into the fast-dispersing solid dosage form maybe in the form of an extract, a purified allergen, a modified allergen,a recombinant allergen or a mutant of a recombinant allergen. Anallergenic extract may naturally contain one or more isoforms of thesame allergen, whereas a recombinant allergen typically only representsone isoform of an allergen. In a preferred embodiment the allergen is inthe form of an extract. In another preferred embodiment the allergen isa recombinant allergen. In a further preferred embodiment the allergenis a naturally occurring low IgE-binding mutant or a recombinant lowIgE-binding mutant.

Allergens may be present in equi-molar amounts or the ratio of theallergens present may vary preferably up to 1:20.

In a further embodiment of the invention the low IgE binding allergen isan allergen according to WO 99/47680 or WO 02/40676 or PCT/DK03/00322(“Allergen mutants”).

Several laboratory tests are available for characterising an allergen.The most widely used techniques are sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing(IEF), crossed immunoelectrophoresis (CIE) and Rocket ImmunoElectrophoresis (RIE). The quantification of individual allergens may beperformed by a variety of quantitative immunoelectrophoretic techniques(QIE), Radial Immune Diffusion (RIE) or by enzyme-linked immunosorbentassays (ELISA). The determination of total allergenic activity is mostfrequently performed by radio allergosorbent test (RAST), Magic Liteassay (LIA) or related techniques. ELISA-based techniques may also beused.

Guidance to the normally applied, acceptable tests measuring biopotencyare found e.g. in Note for Guidance on Allergen Product; The EuropeanAgency for the Evaluation of Medicinal Product, CPMP_BWP_243_96, London,1996.

The classification of an allergen as a major allergen can be subject toseveral tests. An allergen is commonly classified as a major allergen ifat least 25% of the patients shows strong IgE binding (score 3) and atleast moderate binding (score 2) from 50% of the patients, the bindingbeing determined by an CRIE (Crossed Radio Immune Electrophoresis) (CRIEStrong binding, i.e. visible IgE-binding on an X-ray film after one day;CRIE Moderate binding, i.e. binding after 3 days; CRIE Weak binding,i.e. binding after 10 days). Strong IgE binding from at least 10% of thepatients classifies the allergen as an Intermediate allergen and clearlyspecific binding from less than 10% of the patients classifies it as aMinor allergen. Other methods may also be used in determining the IgEbinding of for instance IgE-blots.

In classical incremental dosage desensitisation, where the dose ofallergen in the form of a fast-dispersing solid dosage form is increasedto a certain maximum, the preferred potency of a unit dose of the dosageform is from 150-1,000,000 SQ-U/dosage form, more preferred the potencyis from 500-500,000 SQ-U/dosage form, more preferred the potency is from500-375,000 SQ-U/dosage form, more preferably the potency is from2500-375,000 SQ-U/dosage form, more preferred the potency is from2500-250,000 SQ-U/dosage form, more preferred 25,000-250,000 SQ-U/dosageform, more preferably 25,000-125,000 SQ-U/dosage form, more preferred25,000-100,000 SQ-U/dosage form and most preferable 25,000-75,000SQ-U/dosage form.

In another embodiment of the invention the solid dosage form is arepeated mono-dose, preferably within the range of from 2500-375,000SQ-U/dosage form, more preferably 2500-250,000 SQ-U/dosage form, morepreferably 25,000-250,000 SQ-U/dosage form, more preferably from25,000-125,000 SQ-U/dosage, even more preferably from 25,000-100,000SQ-U/dosage form, and most preferably from 25,000-75,000 SQ-U/dosageform.

In a particular preferred embodiment the solid dosage from comprises angrass allergen extract wherein the potency is from 150-1,000,000SQ-U/dosage form, more preferred the potency is from 500-500,000SQ-U/dosage form, more preferred the potency is from 500-375,000SQ-U/dosage form, more preferably the potency is from 2500-375,000SQ-U/dosage form, more preferred the potency is from 2500-250,000SQ-U/dosage form, more preferred 25,000-250,000 SQ-U/dosage form, morepreferably 25,000-125,000 SQ-U/dosage form, more preferred25,000-100,000 SQ-U/dosage form and most preferable 25,000-75,000SQ-U/dosage form.

In yet a further embodiment the potency of a solid dosage form accordingto the invention is from about 5-50,000 BAU/dosage form, more preferredthe potency is from 15-25,000 BAU/dosage form, more preferably thepotency is from about 15-17,600 BAU/dosage form, more preferably thepotency is from about 65-17,600 BAU/dosage form, more preferably thepotency is from about 65-15,000 BAU/dosage form, more preferably thepotency is from about 650-15,000 BAU/dosage form, more preferred650-6,000 BAU/dosage form, more preferred 650-4,700 BAU/dosage form,most preferable 650-3,500 BAU/dosage form.

In another embodiment of the invention the solid dosage form is arepeated mono-dose, preferably within the range of about 65-17,600BAU/dosage form, more preferably about 65-15,000 BAU/dosage form, morepreferably about 650-15,000 BAU/dosage form, more preferred 650-6,000BAU/dosage form, even more preferred 650-4,700 BAU/dosage form, mostpreferable 650-3,500 BAU/dosage form.

In a particular preferred embodiment the solid dosage from comprises angrass allergen extract wherein the potency is from about 5-50,000BAU/dosage form, more preferred the potency is from 15-25,000 BAU/dosageform, more preferably the potency is from about 15-17,600 BAU/dosageform, more preferably the potency is from about 65-17,600 BAU/dosageform, more preferably the potency is from about 65-15,000 BAU/dosageform, more preferably the potency is from about 650-15,000 BAU/dosageform, more preferred 650-6,000 BAU/dosage form, more preferred 650-4,700BAU/dosage form, most preferable 650-3,500 BAU/dosage form

1 mg allergen extract normally contains between 100,000 and 1,000,000SQ-unit. This means that 1,000,000 SQ are contained in from 1 mg extractto 10 mg allergen extract, and that 100,000 SQ are contained in from 0.1mg extract to 1 mg allergen extract. In a similar manner, any SQ dosemay be transformed into an allergen extract dose range. On this basis,the above dose ranges given in SQ may be recalculated into dose rangesin mg or μg allergen extract, wherein for the lower SQ limit of a range,the lower limit of the corresponding allergen extract range is used, andwherein for the upper SQ limit of a range, the upper limit of thecorresponding allergen extract range is used.

Thus, in a further embodiment a solid dosage form according to theinvention has an allergen extract content of about 0.15 μg-10 mg/dosageform, more preferred an allergen extract content of about 0.5 μg-5mg/dosage form, more preferably an allergen extract content of about 0.5μg-3.75 mg/dosage form, more preferably an allergen extract content ofabout 2.5 μg-3.75 mg/dosage form, more preferably an allergen extractcontent of about 2.5 μg-2.5 mg/dosage form, more preferably an allergenextract content of about 25 μg-2.5 mg/dosage form, more preferred about25 μg-1.25 mg/dosage form, even more preferred about 25 μg-1 mg/dosageform, most preferable about 25 μg-0.75 mg/dosage form.

In another embodiment of the invention the solid dosage form is arepeated mono-dose, preferably within the range of about 2.5 μg-3.75mg/dosage form, more preferably 2.5 μg-2.5 mg/dosage form, morepreferably of about 25 μg-2.5 mg/dosage form, more preferred of about1.5 μg-1.25 mg/dosage form, even more of about preferred 25 μg-1mg/dosage form, most preferable of about 25 μg-0.75 mg/dosage form.

In a further embodiment, a solid dosage form according to the inventionhas a major allergen content of about 0.015 μg-1 mg/dosage form, morepreferred of about 0.05 μg-500 μg/dosage form, more preferably of about0.05 μg-375 μg/dosage form, more preferably of about 0.25 μg-375μg/dosage form, more preferably of about 0.25 μg-250 μg/dosage form,more preferably of about 2.5 μg-250 μg/dosage form, more preferred about2.5 μg-125 μg/dosage form, even more preferred about 2.5 μg-100μg/dosage form, most preferable about 2.5 μg-75 μg/dosage form.

In another embodiment of the invention the solid dosage form is arepeated mono-dose, preferably within the range of 0.25 μg-375 μg/dosageform, more preferably of about 0.25 μg-250 μg/dosage form, morepreferably of about 2.5 μg-250 μg/dosage form, more preferred about 2.5μg-125 μg/dosage form, even more preferred about 2.5 μg-100 μg/dosageform, most preferable about 2.5 μg-75 μg/dosage form.

The content of major allergens may be accounted for by several majorallergens depending on the allergen source in question. Normally thenumber of major allergens is in the range of 1-10, mostly 1-5.

The major allergen may be comprised in an allergen extract or berecombinantly produced. Recombinant major allergens may be used in thesame amount as in allergen extracts comprising such major allergen or inhigher doses. Higher doses are believed to be more effective, but arealso believed to be associated with a risk of potentially more frequentor more severe side effects.

In a further preferred embodiment major allergens include grass group 1allergen e.g. phl p 1, lol p 1, sor h 1, dac g 1, cyn d 1, hol l 1, phaa 1, grass group 2/3 allergen e.g. phl p 2/3, lol p 2/3, grass group 5allergen e.g. phl p 5, lol p 5, dac g 5, poa p 5, grass group 6 allergene.g. phl p 6, poa p 6, tree pollen group 1 allergen e.g. bet v1, aln g1, cor a 1, car b 1, mite group 1 allergen e.g. der p 1, der f 1, eur m1, mite group 2 allergen e.g. der p 2, der f 2, eur m2, cat allergene.g. fel d 1, cedar group 1 and group 2 allergen e.g. cry j 1, cry j 2,short or giant ragweed pollen allergen e.g. amb a 1, amb a 2, amb 1, ambt 2.

A dose response effect is found cf. example 6 for solid allergen dosageforms administered oromucosally. As allergic individuals are aninhomogeneous group displaying different symptoms and different degreesof severity of their symptoms when exposed to even the same allergen,effective doses may vary. Some patients can tolerate larger doseswithout experiencing unacceptable side effects, while others arehypersensitive. In some cases escalating doses may be given to reachhigh dose levels as it is generally believed that larger doses arebelieved to be more effective doses. It is believed that for themajority of the average allergic population an effective dose ofallergen according to the invention will preferably be between 65BAU/solid dosage form-17,600 BAU/solid dosage form, but a dose as low as4 and up to 47,000 BAU may be applicable for other allergic patients.Equally hereto, a dose of allergen extract of 0.5 μg-3.75 mg/dosageform, or a dose with a major allergen content of 0.05 μg-375 μg/dosageform may be suitable for an average allergic individual.

For hypoallergenic variants of major allergens, i.e. allergens with adecreased ability to caused immediate or late phase allergic reactions,a dosage form according to the invention preferably contains 10-100times more major allergen per dosage form. Such hypoallergenic variantsmay be of recombinant or natural origin.

The allergen content of a solid dosage form according to the inventioncan be determined by routine immune assays such as CIE (Cross ImmuneElectrophoresis), RIE (Radio Immune Electrophoresis) and SDS-PAGE(Sodium Dodecyl Sulphate Poly Acrylamide Gel Electrophoresis) and immuneassays such as ELISA and Magic Like Specific IgE assay (LIA) againstextract components such as major allergens.

In order to ensure a sufficient shelf life for the final product, dosageforms which do not significantly change after manufacture with respectto physical and chemical properties, e.g. potency and content of theallergen, mechanical robustness and organoleptical properties, arepreferred.

For the purpose of this invention the stability of the activeingredient, i.e. the allergen, is assessed by means of major allergencontent. Further to that stability is preferably also assessed by meansof potency measurements of the allergen such as total allergen activity.

The “initial allergenic activity” or the “initial content of at leastone major allergen” of a solid dosage form means the value of the“allergenic activity” or the “content of at least one major allergen” inthe final dosage form after the manufacturing process has ended.

The “theoretical allergenic activity” or the “theoretical content of atleast one major allergen” of a solid dosage form means the value of the“allergenic activity” or the “content of at least one major allergen” ofa dose of the added allergen, e.g. in the form of an extract, beforebeing formulated into a solid dosage form.

Loss in the allergen content of at least one major allergen ispreferably less than 50% of the total initial content, more preferablyless than 30% of the total initial content, more preferably less than20% of the total initial content, more preferably less than 15% of theinitial content, more preferably less than 10% of the initial content,more preferably less than 5% of the initial content, more preferablyless than 2% of the initial content.

Further loss in total allergen activity according the method describedin Example 1 should preferably be less than 50% of the total initialactivity, more preferably less than 30% of the total initial activity,more preferably less than 20% of the total initial activity, morepreferably less than 15% of the total initial activity.

Normally, stability testing is conducted according to the currentguidelines by ICH (e.g. ICH guideline ICQ/Q1AR2 (Adopted by CPMP, March2003, issued as CPMP/ICH/2736/99)) and FDA. Conditions for stabilitytesting are often referred to as zone 1-4 conditions. Zone 1 and 2represent climatic conditions in EC, Japan and US. Normally, soliddosage forms according to the present invention in their final containershould preferable be stable for a least 3 months, more preferably for atleast 6 months, more preferably for at least 12 months, even morepreferred for at least 18 months, most preferred at least for 2 years at“long term” conditions 25° C./60 RH, more preferable at intermediatecondition 30° C./65 RH, even more preferable at “accelerated conditions”40° C./75 RH.

To ensure that the solid dosage form is sufficiently robust duringstorage and when handled by the patient, the dosage form needs to have acertain resistance to external force, but at the same time the soliddosage form needs to disintegrate quickly in the mouth.

Thus, further to the stability of the active ingredient i.e. theallergen solid dosage forms can also be assessed by additionalparameters, such as mechanical robustness like friability, tensilestrength, and peak load to fracture. In addition, the stability of thesolid dosage forms can be assessed by physical properties such as thedispersion time and organoleptical properties like visual appearance ofthe dosage form.

These parameters can be evaluated by e.g. measurements of Peak load tofracture or tensile strength of the solid dosage forms of the currentinvention. As it is apparent from the equation from which the tensilestrength can be calculated, the tensile strength value obtained dependson a number of parameter, which are subject to variation. For example,thickness or the diameter of the solid dosage form will contribute tothe variation of value. Therefore, Peak load to fracture is believed tobe an even more accurate parameter for evaluation of the robustness ofthe solid dosage units of the current invention.

In an embodiment the solid dosage form has a Peak load to Fracture notless than 0.05 Kgf and below 0.9 KgF. Preferred are dosage forms whichhave a Peak load to Fracture which is between 0.05-0.9 KgF, morepreferred between, 0.1-0.8 KgF, most preferred between 0.1-0.6 KgF.

In a further embodiment of the current invention the solid dosage formhas a tensile strength less than 1.0 N/mm2, more preferred below 0.9N/mm2.

Preferably fast-dispersing dosage form disintegrates instantaneously orquickly in the mouth upon contact with the saliva in order to ensuremaximum exposure of allergen to immune competent tissue of the mucosabefore swallowing. In a preferred embodiment the solid dosage formdisintegrates in less than about 90 seconds, preferably in less thanabout 60 seconds, preferably in less than about 30 seconds, morepreferably in less than about 20, more preferably in less than about 15seconds, even more preferably in less than about 10 seconds in the oralcavity, even more preferably in less than about 5 seconds, mostpreferably in less than about 2 seconds in the oral cavity.

In a preferred embodiment of the invention, the compositions of theinvention are fast-dispersing solid dosage forms comprising a solidnetwork of the allergen and any water-soluble or water-dispersiblematrix. The network is obtained by subliming solvent from a compositionin the solid state, the composition comprising a solution of theallergen and the matrix. More preferably the network is obtained bylyophilization.

Pharmaceutically acceptable excipients forming part of the matrix in thefast-dispersing solid dosage form according to invention are matrixforming agents and additionally other suitable excipients such asantacids, diluents, enhancers, mucoadhesive agents, flavouring agents,taste masking agents, preservatives, antioxidants, surfactants,viscosity enhancers, colouring agents, pH modifiers, sweeteners etc.These excipients are all selected in accordance with conventionalpharmaceutical practice in a manner understood by persons skilled in theart of formulating allergen therapeutics.

Matrix forming agents suitable for use according to the presentinvention include excipients derived from animal or vegetable proteinssuch as gelatines, dextrins and soy, wheat and psyllium seed proteins;gums such as acacia, guar, agar and xanthan; polysaccharides; starch andmodified starch, alignates; carboxymethylcellulose; carrageenans;dextrans; pectins; synthetic polymers such as polyvinylpyrrolidone; andpolypeptide/protein or polysaccharide complexes such as gelatine-acaciacomplexes. Gelatines are a heterogeneous mixture of water solublecolloid macromolecules. Such heterogeneous mixtures of average molecularweights distribution may be obtained from hydrolytic action on collagenrich material of animal origin such as bone, skin, tendons, ligamentsetc. Gelatines may be derived from mammal e.g. cattle, pig ornon-mammals e.g. warm or cold-water fish. Gelatines can be hydrolysed ornon-hydrolysed, cross-linked or non-cross-linked. They can further be ofa gelling or non-gelling type, the non-gelling type typically beingderived from coldwater fish. In another particular embodiment starch isused. Starches are complex mixtures of carbohydrate polymers.

Other matrix forming agents suitable for use according to the presentinvention include sugars such as mannitol, dextrose, lactose, galactoseand trehalose; cyclic sugars such as cyclodextrin; inorganic salts suchas sodium phosphate, sodium chloride and aluminium silicates; and aminoacids having from 2 to 12 carbon atoms such as a glycine, L-alanine,L-aspartic acid, L-glutamic acid, L-hydroxyproline, L-isoleucine,L-leucine and L-phenylalanine.

The solid dosage form preferably comprises at least about 50% W/W of atleast one matrix forming agent of the dosing solution. The term “dosingsolution” as used in this context means a non-solid volume of aformulation of the matrix forming agents, the allergen and otheroptional excipients that is prepared before the solidification step.

In one embodiment of the invention the dosing solution for forming thesolid dosage form comprises about 5-30% W/W, more preferably about 5-20%W/W, even more preferred between about 5-12% W/W of at least one matrixforming agent.

The need for dry matter content of the dosing solution will also dependon the dimensions of the tablet. Preferably the solid dosage formsaccording to the present invention have a diameter between about 3 toabout 30 mm, more preferably between about 5 to about 20 mm. Preferablythe solid dosage forms according to the present invention have a weightbetween about 1 to about 100 mg, more preferably between about 10 toabout 50 mg, most preferably between about 25 to about 35 mg. Preferablythe solid dosage forms according to the invention have a height betweenabout 0.5 to about 7.5 mm, more preferably between about 1 to about 5mm.

A fast-dispersing solid dosage form comprising fish gelatine andmannitol as matrix-forming excipients has been found to be especiallyadvantageous with respect to stability, visual appearance, lowfriability, tensile strength, peak load to fracture and mouth feel. In apreferred embodiment the fast-dispersing solid dosage forms comprises asolid network of the allergen and matrix form agents in the form of fishgelatine and mannitol. In order to obtain a solid network, the ratio offish gelatine to mannitol should be controlled. In a preferredembodiment the ratio of fish gelatine to mannitol is from about 2:20 toabout 20:1, more preferably from about 2:10 to about 10:1, mostpreferably from about 3:5.5 to about 6.5:3.

In yet a further embodiment the ratio of fish gelatine to mannitol is4:3.

In another embodiment the ratio of fish gelatine to mannitol is 6.5:5.5.

In a further embodiment the ratio of fish gelatine to mannitol is6.0:5.08.

The solid dosage form according to the present invention may bemanufactured from a dosing solution, which is first frozen and thenfreeze dried. In a preferred embodiment the content of fish gelatine isbetween about 2-20% W/W of the dosing solution and the content ofmannitol is between about 1-20% W/W of the dosing solution. In anotherpreferred embodiment the content of fish gelatine is between about 2-10%W/W of the dosing solution and the content of mannitol is between about1-10% W/W of the dosing solution. In a further preferred embodiment thecontent of fish gelatine is between about 3-6.5% W/W of the dosingsolution and the mannitol is between about 3-5.5% W/W of the dosingsolution.

In yet a further embodiment the matrix comprises about 4% W/W of thedosing solution fish gelatine and about 3% mannitol W/W of the dosingsolution

In another embodiment the matrix comprises about 6.5% W/W fish gelatineof the dosing solution and about 5.5% W/W mannitol of the dosingsolution.

In a further embodiment the matrix comprises 6.0% W/W fish gelatine ofthe dosing solution and 5.08% W/W mannitol of the dosing solution.

A fast-dispersing solid dosage form comprising starch and mannitol asmatrix-forming excipients has also been found to be especiallyadvantageous with respect to stability, visual appearance, lowfriability, tensile strength, peak load to fracture and mouth feel. In apreferred embodiment the fast-dispersing solid dosage forms comprises asolid network of the allergen and matrix form agents in the form ofstarch preferably pre-gelatinised from e.g. potato, wheat, maize, cornor rice and mannitol. In order to obtain a solid network, the ratio ofstarch to mannitol should be controlled. In a preferred embodiment theratio of starch to mannitol is from about 2:20 to about 20:1, morepreferably from about 2:10 to about 10:1, most preferably from about3:5.5 to about 6.5:3.

In yet a further embodiment the ratio of starch to mannitol is 1:1.

The solid dosage form according to the present invention is manufacturedfrom a dosing solution, which is first frozen and then freeze dried. Ina preferred embodiment the content of starch is between about 2-20% W/Wof the dosing solution and the content of mannitol is between about1-20% W/W of the dosing solution. In another preferred embodiment thecontent of starch is between about 2-10% W/W of the dosing solution andthe content of mannitol is between about 1-10% W/W of the dosingsolution. In yet a further preferred embodiment the content of starch isbetween about 3-6.5% W/W of the dosing solution and the mannitol isbetween about 3-5.5% W/W of the dosing solution

In another embodiment the matrix comprises about 4.4.% W/W starch of thedosing solution and about 4.4% W/W mannitol of the dosing solution.

Preferably pH is adjusted prior to solidification of the allergen andmatrix containing solution to avoid denaturation of the allergen,precipitation and assure a stable product. The optimum pH for differentallergens in solution span almost the entire pH range as does theirisoelectric point (pI). Mixtures of allergens like extracts equally haveoptimum pH for solubility and stability determined by factors like theconcentration of the individual allergens in the extract. Therefore anindividual determination of a feasible range of pH for a formulationaccording to this invention may be envisaged. The optimum pH for theallergen in question is determined by carrying out accelerated stabilitystudies with formulations with different pH. The design of such studiesis known to the person skilled in the art.

Preferably matrix compositions containing an allergen extract should beadjusted to pH between 3.5-10, more preferably 4-9, most preferably 6-9.

Furthermore it is well known in the art that ionic strength may be aparameter affecting the stability of a freeze-dried solid dosage formprimarily through its effect of the freeze-drying processes. Also it isknown that high ionic strengths affect precipitation. Accordingly, anoptimum must be established by measurements well known to one skilled inthe art. Preferably the ionic strength of an extract of 10 μg/ml is inbetween 1-1500 μS/cm (S=Simens), more preferably between 300-800 μS/cm,most preferably about 500 μS/cm, for a matrix and allergen containingsystem it is preferred that the ionic strength is between 1-2000 μS/cm,more preferably about 500-1500 μS/cm.

Solid dosage forms according to the invention may further comprisecolouring agents, flavours, pH modifiers or taste-masking agents.Suitable colouring agents include red, black and yellow iron oxides andFD & C dyes such as FD & C blue No. 2 and FD & C red No. 40. Suitableflavouring agents include mint, raspberry, liquorice, orange, lemon,grapefruit, caramel, vanilla, cherry and grape flavours and combinationof these. Suitable pH modifiers include citric acid, tartaric acid,phosphoric acid, hydrochloric acid and maleic acid. Suitable sweetenersinclude aspartame, acesulfame K and thaumatic. Suitable taste-maskingagents include sodium bicarbonate, ion-exchange resins, cyclodextrininclusion compounds, adsorbates or microencapsulated actives.

Adjuvants are normally used to enhance the absorption of the allergen aswell as to enhance the immune-stimulating properties of the allergen.

In another preferred embodiment of the invention the fast-dispersingsolid dosage form according to the invention does not comprise anadjuvant.

It has also surprisingly been found that it is not necessary toincorporate an adjuvant into the fast-dispersing solid dosage form inorder to enhance the immune-stimulating properties of the allergen inquestion. That is to say a solid dosage from as described herein canraise a specific immune response as demonstrated in example 6.

In one embodiment of the invention at least one adjuvant is incorporatedinto the dosage form according to the invention. Examples of suitableadjuvants are aluminium salts, aluminium hydroxide such as Alhydrogel®,non-toxic bacterial fragments, cytokines, cholera toxin (and detoxifiedfractions thereof), cholera toxin subunit b, chitosan, homologousheat-labile fragments of E. coli (and detoxified fractions thereof),saponins, bacterial products such as lipopoly-saccharides (LPS) andmuramyl dipeptide (MDP), liposomes, CpG (immunostimulatory DNAsequences), lactide/glycolide homo±copolymers in the form ofmicroparticular polymers etc. The use of adjuvants in allergenpharmaceutical product e.g. vaccines are often reasoned by the fact theallergens in question are not able to penetrate the barrier to bepassed. The adjuvants thus may serve as absorption enhancing agents orthey may act as immunostimulants. The use of adjuvants may, however, beassociated with serious draw backs such as unintended stimulation ofvarious mechanisms of the immune response, systemic lupus erythematosusor affecting the barrier capabilities of the mucosal membranes and thusallowing the passage of hazardous substances. Further from an industrialpoint of view addition of an adjuvant further constitute furthermanufacturing and material cost besides the large demand fordocumentation in respect to drug registration.

A non-compressed fast-dispersing solid dosage form may be mucoadhesiveto some extent. However in a preferred embodiment of the invention, itmay be necessary to further add mucoadhesive excipients to said dosageform in order to increase the contact time of the dosage form with themucosa of the oral cavity. Suitable mucoadhesive excipients arepolyacrylic polymers such as carbomer and carbomer derivatives;cellulose derivatives such as hydroxypropylmethylcellulose,hydroxyethylcellulose, hydroxypropylcellulose and sodiumcarboxymethylcellulose; natural polymers such as gelatine, sodiumalginate, pectin and glycerol.

In further embodiment of the invention the allergen dosage formdissolved in saliva is not swallowed until 3 min after administration inorder to allow sufficient contact time for e.g. absorption over themucosal membrane in the mouth.

In yet a further preferred embodiment the allergen dosage form is notdiluted in the oral cavity e.g. by intake of a fluid like water untilafter 5 min.

Adverse events or side effects are known to exist also in connectionwith allergy treatment. In particular treatment aiming at modulating anongoing response in a sensitised individual may pose a risk for inducingside effect upon administration of allergen. Normally side effects seenin connection with oromucosal treatment are reported in the eye, nose,mouth, the upper and lower airway and depending on degree foundacceptable. Most common are itching phenomena. Adverse reactions, suchas anaphylactic shock, swelling of upper and lower airways, difficultybreathing, blood pressure drop, cardiac arrest would not be acceptable.

The fast-dispersing solid dosage form according to the invention may bemanufactured and packed in disposable containers containing amultiplicity of solid dosage forms i.e. multi dosage containers. Themethods and materials as described in U.S. Pat. Nos. 5,729,958 and5,343,762 are particularly favoured. Examples of suitable multi dosagecontainers are All Aluminium Blister packs, blister packs made ofpolymers e.g. polypropylene, blister packs of PVC and blister packsformed from PVC/PVdC laminate and sealed with e.g. aluminium laminatedto calendered kraft paper, Aclar® or Triplex®.

In an embodiment the fast-dispersing dosage form is manufactured andpacked in blister packs formed from PVC/PVdC laminate and sealed withaluminium laminated to calendered kraft paper. In another embodimenthereof the blister pack are enclosed in an aluminium sachet of suitablesize, composed of aluminium laminated to calendered kraft paper.

In yet another embodiment the fast-dispersing dosage form is packed inblister packs formed from aluminium and sealed with aluminium laminatedto calendered kraft paper.

In a further embodiment the fast-dispersing dosage form is packed inmultilamilar blister packs formed from e.g. five layer aluminiumlaminate and sealed with aluminium laminated to calendered kraft paper.

In yet another embodiment the fast-dispersing dosage form is packed inblister packs formed from aluminium laminate and sealed with aluminiumlaminated to calendered kraft paper in such a way that is difficult forchildren to open the blister pack e.g. child resistant packs.

A solid dosage form of this type generally may be characterized by a lowmechanical strength compared to compressed tablets, because of theinherent nature of such a non-compressed dosage form. This may result inthe release of residual particles containing the allergen on removalfrom the blister pocket and during handling of the dosage form by thepatient. In most situations this is of no or mainly cosmetic importance.However, this is especially detrimental when the active ingredient is anallergen, because low amounts allergen can elicit an allergic reactionin a disposed person or sensitise a previously non-sensitizedindividual. Normally exposure is in the range of 10 μg/year to majorallergen protein accumulated for e.g. pollen allergens or dust miteallergens, which is adequate to give sensitisation or symptoms.

Upon handling the solid dosage forms, allergens may come in contact withtarget organs like the airways or the eye and elicit a response in anallergic person. One dosage form may contain as much allergen as aperson is exposed to over one year or more depending upon the nature ofthe exposure. It is possible to induce eye symptoms in allergic patientsusing a conjunctival allergen challenge. Based on such challenge studiesit can be estimated how much allergen extract is needed to induceconjunctival symptoms. In a population of patients with severegrass-pollen induced hayfever, the lowest dose of grass pollen extractcausing conjunctival symptoms was proposed to be 3000 SQ-U/ml×0.05ml=150 SQ-U (median value) (S. R. Durham, S. M. Walker, E. M. Varga, M.R. Jacobson, F. O'Brien, W. Noble, S. J. Till, Q. A. Hamid, and K. T.Nouri-Aria. Long-term clinical efficacy of grass-pollen immunotherapy.N. Engl. J. Med. 341 (7):468-475, 1999).

In order to ensure that allergen containing residues from the soliddosage form is not released to the environment upon opening the multidosage container, it is important that the friability of the dosage formis as low as possible without jeopardising the allergen release from thedosage form following oral administration.

Thus, in one embodiment less than about 500 SQ-U may be released fromeach solid dosage form during manual handling, more preferably less thanabout 250 SQ-U, more preferably less than about 150 SQ-U, morepreferably less than about 75 SQ-U, more preferably less than about 25SQ-U, most preferably less than about 10 SQ-U.

In another embodiment less than about 13 BAU may be released from eachsolid dosage form during manual handling, more preferably less thanabout 7 BAU, most preferably less than about 5 BAU, more preferably lessthan about 1.95 BAU, more preferably less than about 0.65 BAU, and mostpreferably less than about 0.26 BAU.

In a further embodiment less than about 0.5 μg allergen extract may bereleased from each solid dosage form during manual handling, morepreferably less than about 0.25 μg allergen extract, most preferablyless than about 0.15 μg allergen extract, more preferably less thanabout 0.075 μg allergen extract, more preferably less than about 0.025μg allergen extract, and most preferably less than about 0.01 μgallergen extract.

In yet another embodiment less than about 0.05 μg major allergen may bereleased from each solid dosage form during manual handling, morepreferably less than about 0.025 μg major allergen, most preferably lessthan about 0.015 μg major allergen, more preferably less than about0.0075 μg major allergen, more preferably less than about 0.0025 μgmajor allergen, most preferably less than about 0.001 μg major allergen.

In a preferred embodiment of the present invention the residual contentof dust in the multi dosage container after removing the dosage form donot exceed about 2% of total allergen content, more preferred about 0.5%of total allergen content of a solid dosage form and more preferablyabout 0.2% of total allergen content of a solid dosage form and mostpreferably about 0.1% of total allergen content of a solid dosage form,more preferably about 0.01%, more preferred about 0.005%, more preferredabout 0.003% of total allergen content of a solid dosage form, mostpreferred about 0.001% of the total allergen content of a solid dosageform.

As used herein “friability test” shall refer to any suitable test thatmeasures the ease with which a solid dosage form crumbles, falls topieces, are reduced to powder. Suitable friability test to use in thecurrent invention are illustrated below and include EuropeanPharmacopoeia 3rd edition (EP 3rd ed.) Pharmaceutical technicalprocedures 2.9.7. Normally friability testing of tablets is performed asset out in the EP 3rd ed. 2.9.7. and USP <1216>, wherein loss of weightis assessed as a parameter of an intact dosage form. The EP 3rd ed.2.9.7. friability test uses a drum having a diameter of 286 mm and about39 mm in depth. A sample of the tablets are placed on a sieve no. 100and any loose residuals are removed by use of air pressure or a softbrush. The tablets are weighed and thereafter placed in the drum. Thetablets are rotated for 100 times in the drum. Loose residuals are thenremoved as just described and the tablets are weighed again. The resultis then expressed as the loss of mass and is calculated as a percentageof the initial mass. According to USP <1216> a drum with a diameterbetween 283 and 291 and a depth between 36 and 40 mm may be used and arotation of 25±1 rpm. Accordingly, the intactness of the current dosageform may be assessed by visual inspection and measurement of tabletweight upon having been subject to such a method. Alternatively, due tothe low weight of dosage forms according to the invention the weighingcan be replaced with an immune assay specific for the allergen inquestion.

The use of a modified friability test has been found to be a useful toolin assessing which compositions are most stable with respect torobustness and mechanical strength.

In an embodiment the friability of said solid dosage form measured asthe amount of allergen released is less than about 500 SQ-Upper soliddosage form, more preferably less than about 250 SQ-Upper solid dosageform, more preferably less than about 150 SQ-Upper solid dosage form,more preferably less than about 75 SQ-Upper solid dosage form, morepreferably less than about 50 SQ-Upper solid dosage form, morepreferably less than about 25 SQ-Upper solid dosage form, mostpreferably less than about 10 SQ-Upper solid dosage form in any suitablefriability test that exerts a sufficient external force on thecompositions to be tested.

In a more preferred embodiment the friability measured as the amount ofallergen released is less than about 500 SQ-Upper solid dosage form,more preferably less than about 250 SQ-Upper solid dosage form, morepreferably less than about 150 SQ-Upper solid dosage form, morepreferably less than about 75 SQ-Upper solid dosage form, morepreferably less than about 50 SQ-Upper solid dosage form, and morepreferably less than about 25 SQ-Upper solid dosage form, mostpreferably less than about 10 SQ-Upper solid dosage form in a friabilitytest performed according to the Pharmacopoeia EP 3^(rd) ed.

In an even more preferred embodiment the friability measured as theamount of allergen released is less than 500 SQ-Upper solid dosage form,more preferably less than 250 SQ-Upper solid dosage form, morepreferably less than about 150 SQ-Upper solid dosage form, morepreferably less than about 75 SQ-Upper solid dosage form, morepreferably less than about 50 SQ-Upper solid dosage form, and morepreferably less than about 25 SQ-Upper solid dosage form, mostpreferably less than about 10 SQ-Upper solid dosage form in an methodcomprising the following steps;

-   -   (a) placing individual sealed blisters each containing a solid        dosage form in equipment suitable for friability measurements;    -   (b) moving the sealed blister containing the solid dosage form        for an appropriate time and at an appropriate velocity;    -   (c) removing the sealed blister containing the solid dosage        form;    -   (d) opening the blister and placing the solid dosage form and        any residues in a container;    -   (e) removing the solid dosage form unit from the container        leaving any loose residuals in said container;    -   (f) performing an allergen specific assay on said residues        determining the allergen content in said residues; and    -   (g) optionally calculating the percentage allergen content in        said residues of the total allergen content of the solid dosage        form unit.

In another embodiment the friability of said solid dosage form measuredas the amount of allergen released is less about 0.5 μg allergen extractper solid dosage form during manual handling, more preferably less thanabout 0.25 μg allergen extract per solid dosage form, more preferablyless than about 0.15 μg allergen extract per solid dosage form, morepreferably less than about 0.075 μg allergen extract per solid dosageform, more preferably less than about 0.025 μg allergen extract persolid dosage form, most preferably less than about 0.01 μg allergenextract per solid dosage form in an method comprising the followingsteps;

-   -   (a) placing individual sealed blisters each containing a solid        dosage form in equipment suitable for friability measurements;    -   (b) moving the sealed blister containing the solid dosage form        for an appropriate time and at an appropriate velocity;    -   (c) removing the sealed blister containing the solid dosage        form;    -   (d) opening the blister and placing the solid dosage form and        any residues in a container;    -   (e) removing the solid dosage form unit from the container        leaving any loose residuals in said container;    -   (f) performing an allergen specific assay on said residues        determining the allergen content in said residues; and    -   (g) optionally calculating the percentage allergen extract        content in said residues of the total allergen extract content        of the solid dosage form unit.

In yet another embodiment the friability measured as the amount ofallergen released is less than about 0.05 μg major allergen, morepreferably less than about 0.025 μg major allergen, more preferably lessthan about 0.015 μg major allergen per solid dosage form, morepreferably less than about 0.0075 μg major allergen per solid dosageform, more preferably less than about 0.0025 μg major allergen per soliddosage form, most preferably less than about 0.001 μg major allergen persolid dosage form in an method comprising the following steps;

-   -   (a) placing individual sealed blisters each containing a solid        dosage form in equipment suitable for friability measurements;    -   (b) moving the sealed blister containing the solid dosage form        for an appropriate time and at an appropriate velocity;    -   (c) removing the sealed blister containing the solid dosage        form;    -   (d) opening the blister and placing the solid dosage form and        any residues in a container;    -   (e) removing the solid dosage form unit from the container        leaving any loose residuals in said container;    -   (f) performing an allergen specific assay on said residues        determining the major allergen content of at least one major        allergen in said residues; and    -   (g) optionally calculating the percentage of at least said major        allergen in said residues of the total major allergen content of        the solid dosage form unit.

In another preferred embodiment of the method

between 1 and 100 blisters containing the solid dosage form an equipmentfor friability measurements as described in European PharmacopoeiaV.2.9.7 is used in step a), the solid dosage forms are rotated for 100turns at 25±1 rpm in step b), and the allergen specific assay is animmunochemical allergen specific assay in step f).

In a further preferred embodiment of the method for measuring friabilitythe allergen content is determined by an ELISA assay.

Furthermore, the oral dosage form must have an appealing appearance.Hence, as a part of the quality control the fast-dispersing solid dosageforms according to the invention are preferably subjected to visualinspection e.g. colour, shape, irregularities and defects.

In order to ensure optimum compliance of the patient, the dosage formmay also be tested for mouth feel. For example, the patients perceivethe dosage form as being pleasant when it is placed in the mouth andallowed to disintegrate.

As allergens are very bio-potent for the allergic person i.e. even smallamount may trigger a response, uniformity of content is an importantparameter during treatment to ensure, for example, that a patternexperienced by a patient is reproducible when taking the same dose.Preferably the variation of content of allergen of units within a multidosage container is within ±10%, preferably within ±7%, most preferablewithin ±5% compared to the set dose.

A multi dosage container may contain any conceivable number offast-dispersing solid dosage forms. Preferably solid allergen dosageforms are packaged and used as a group. Individual solid dosage formsare packaged by dispensing as a liquid mixture into individualcontainers, followed by removal of water. Multiple blisters may bearranged in larger sheets and multiple sheets may be packaged and soldtogether. For example, a container such as a blister pack may comprise amultiplicity of solid dosage forms preferably 1-100 solid dosage forms,more preferably 1-35, and most preferably 1-10 solid dosage forms perblister pack. Another embodiment provides economy of manufacture,distribution and storage via the use of multiple blister packs that may,for example provided 2, 3, 4, 5, 6 or more for a single intended courseof treatment use. This particular packaging scheme particularly isuseful for mono-dose treatments, and is made possible by dosingregimens, as described herein, that lack an up-dosing step.

Another embodiment provides a treatment pack for the treatment ofallergy or alleviating symptoms of allergy. The treatment pack comprisesa sealed package of multiple solid dosage forms, each of which comprisesan effective amount of an allergen. The treatment pack may contain forexample, at least 2, 4, 6, 7, 10, 14, 30, 60, 90, 100, 120, 200, 240, ormore solid dosage forms. The treatment pack may contain enough dosageforms for an entire treatment, or enough for a portion of a treatment.Advantageously the treatment pack contains at least one month's supplyof unit doses.

In another advantageous embodiment of the invention, all solid dosageforms of a treatment pack contain the same allergen dosage, thusremoving the necessity for manufacturing, distributing, and storingmultiple dosage units for treatment of a single allergy in anindividual.

A kit comprising a treatment pack may further comprise instructionsconcerning use of the solid dosage form. The instructions may comprise awarning, dosage regimen instruction, or any other information of valueto the user. The instructions physically may comprise, for example, aseparate instruction pamphlet, a paper, a placard, and/or one or morenotices printed on a container such as a box that holds the unit doses.The instructions also may be supplied as a compact disk or othercomputer readable medium, or a video cassette.

Most desirably, the multiple solid dosage forms of a treatment pack havebeen formed from a liquid mixture by an in situ process that removes asuspension and/or solvation liquid, which may comprise water and/orother organic solvents and are fast dispersing.

In another embodiment each of the solid dosage forms of a treatment packare located in individually sealed blisters in a multiple blister pack.

In a further embodiment the solid dosage forms of a treatment packcomprise gelatine, more preferably fish gelatine.

In yet a further embodiment the solid dosage forms of a treatment packfurther comprise mannitol.

In yet another embodiment of a treatment pack the effective amount ofsolid dosage form is between about 2.5 μg-about 3.75 mg extract/soliddosage form.

Clinical allergy manifestation and symptoms are several and may varydepending on the sensitized individual and the allergy inflicted. Commonare symptoms like edema, itching, redness and running of the eyes andnose (rhinitis and conjunctivitis) and symptoms from upper and lowerairway like wheezing, coughing, shortness of breath, skin condition likeeczema, urticaria and itching. Other symptoms like fatigue are alsoexperienced. Symptomatic treatment aims at reducing or affectingseverity of the symptoms or reducing the need for other drugs given inparallel. Symptomatic drug includes antihistamines like H₁ and H₂receptor antagonists, intranasal and systemic corticosteroids,non-steroid anti-inflammatory drugs, nasal decongestants likeadrenoceptor agonists. Treatment and relief of one or more allergicsymptom or the reduction in the need for other medication is a furtherobject of this invention.

Allergy is a widespread disease in mammal including humans and animalssuch as dogs and horses. Thus, a further object of the invention is toprovide a method for treatment of allergy or alleviating symptoms ofallergy in mammals comprising oromucosal administration of an effectiveamount of an allergen vaccine dosage form comprising (a) a matrix, and(b) at least one allergen in any of the above described embodiments.

Treatment of in particular seasonal allergies such as hay fever isnormally associated a particular time of year were exposure to theoffending allergen is present or elevated. The allergen season is willvary with the allergen source e.g. the pollen and the climaticconditions for the allergen source in the particular territory. Thus,the season for an allergen will differ in one part of the world fromanother part of the world depending on the climate, but will normallyfall within the same period of the year for the same territory varyingwith the actual conditions of that year (see for instance “Aerobiologyand inhalant allergies”, Chapter 19, T. A. E. Platts-Mills & W. R.Solomon (Ed. S. Manning) 1993, Mosby-Year Book, St. Louis). This will bewell known to a skilled person when a season is normally expected tostart for a particular allergen in a particular region.

In one embodiment of the invention a methods of treatment is providedincluding a pre-seasonal treatment i.e. an administration of soliddosage forms according to the invention before the allergen season. In aparticular preferred embodiment the pre-seasonal treatment periodcomprises administration of solid dosage according to the invention fora period of more than 2 weeks prior to the allergen season, morepreferably between 4-20 weeks, most preferably between 8-12 weeks.

Another object of the invention is to provide a method for treatment ofallergy or allergic symptoms comprising oromucosal administration of aneffective amount of an allergen vaccine dosage form comprising (a) amatrix, and (b) at least one allergen further comprising at least oneanti-allergic drug e.g. antihistamines and histamine synthesisinhibitors in any of the above described embodiments. Preferably suchanti-allergic drugs include bromapheniramine, cetirizine, fexofenadine,cyproheptadine, dexchlorpheniramine, hydroxizine, ketofene, mequitazine,oxotomide, mizolastine, ebastine, astemizole, carbinoxamide,alimemazine, buclizine, cyclizine, hydrochlorate, doxylamine,tritoqualine.

Also the invention also includes use of a pharmaceutical productcomprising a non-compressed, fast dispersing solid dosage formcomprising an allergen and at least one matrix forming agent fororomucosal treatment of allergy or alleviation of allergy symptoms.

In yet another embodiment of the invention a pharmaceutical productcomprising a stable, fast-dispersing, non-compressed allergen soliddosage form with low friability comprising (a) a matrix, and (b) atleast one allergen further comprising an antihistamine is used fororomucosal treatment of allergy or allergic symptoms.

Another embodiment provides a method for treating allergy or alleviatingsymptoms of allergy, comprising providing a a) treatment pack and b)repeatedly administering oromucosally one or more dosage forms of fromthe treatment pack until symptoms are alleviated.

Also a further embodiment provides a treatment regimen that utilizes asingle dose for completing an entire sensitization treatment, withouthaving to updose i.e. increase with different level(s) of allergen tillreaching the particular dose. This embodiment is advantageous because itsimplifies and economizes on dosage form manufacture, distribution andstorage by not requiring multiple dose quantities for a singletreatment. Moreover, by simplifying a treatment course, patientcompliance is improved, which directly leads to greater clinicaleffectiveness.

Another embodiment of the invention is pharmaceutical product comprisingan orally administerable solid dosage form comprising a matrix formed ofat least one pharmaceutically acceptable material, an effective amountof an allergen for desensitizing a human to said allergen, said dosageform having an allergen content at least about 50% of the initialallergen content after being held for 3 months at 25° C. and 60%relative humidity. Preferably said pharmaceutical product is in form ofa lozenge, tablet, a capsule or a caplet.

The fast-dispersing solid dosage form according to the invention can beprepared by a sublimation process according to the process disclosed inU.S. Pat. No. 4,371,516. Accordingly, a solidified solution of theallergen and the matrix forming excipients is subjected to sublimation.The sublimation process is preferably carried out by freeze-drying thesolution. The solution is contained in a depression of the multi dosagecontainer during the freeze-drying step to produce a solid form in anydesired shape. The multi dosage container can be cooled using liquidnitrogen or solid carbon dioxide. After the freezing step the frozensolution in the multi dosage container is subjected to reduced pressureand, if desired, controlled application of heat to aid the sublimationof the solvent.

The invention further includes the use of an allergen for themanufacture of a stable, fast-dispersing, non-compressed allergenvaccine solid dosage form with low friability comprising (a) a matrix,and (b) at least one allergen form for use in the treatment of allergyor alleviating symptoms of allergy.

In another embodiment a method is provided of producing afast-dispersing, non-compressed solid and stable dosage form suitablefor oromucosal administration having low friability comprising at leastone matrix forming agent and an effective dose for desensitizing anindividual to at least one allergen, comprising the steps of

-   -   (a) preparing an aqueous solution comprising said at least one        allergen and at least one matrix forming agent,    -   (b) introducing the solution into one or more depressions in a        mould    -   (c) subjecting the loaded sheet to freezing and freeze-drying        using standard conditions of shelf temperature and chamber        pressure to obtain said solid dosage form in each depression.

In yet another embodiments a method is provided of producing afast-dispersing, non-compressed solid and stable dosage form suitablefor oromucosal administration having low friability comprising at leastone matrix forming agent and an effective dose for desensitizing anindividual to at least one allergen, comprising the steps of

-   -   (a) preparing an aqueous solution comprising said at least one        allergen and at least one matrix forming agent,    -   (b) introducing the solution into depressions in a laminated        blister sheet    -   (c) subjecting the loaded sheet to freezing and freeze-drying        using standard conditions of shelf temperature and chamber        pressure to obtain said solid dosage form in each depression.

In a preferred embodiment of the method the blister sheet is an allaluminium blister sheet. In a particularly preferred embodiment of themethod, the blister sheet is a multilayered, all aluminium blistersheet.

In yet another embodiment a method is provided for obtaining apharmaceutical product comprising a fast-dispersing, non-compressedsolid and stable dosage form suitable for oromucosal administrationcomprising at least one matrix forming agent and an effective dose fordesensitizing an individual to at least one allergen, said dosage formhaving low friability comprising

1) producing a stable fast-dispersing, non-compressed solid dosage form,

2) measuring the friability of said dosage form in an assay comprisingthe steps of,

-   -   (a) placing individual sealed blisters each containing a solid        dosage form in equipment suitable for friability measurements;    -   (b) moving the sealed blister containing the solid dosage form        for an appropriate time and at an appropriate velocity;    -   (c) removing the sealed blister containing the solid dosage        form;    -   (d) opening the blister and placing the solid dosage form and        any residues in a container;    -   (e) removing the solid dosage form unit from the container        leaving any loose residuals in said container;    -   (f) performing an allergen specific assay on said residues        determining the major allergen content of at least one major        allergen in said residues; and    -   (g) calculating the percentage of at least said major allergen        in said residues of the total major allergen content of the        solid dosage form unit.

detecting whether the dosage form fulfils the requirements for lowfriability, and

3) repeating 1) and 2) until the requirements for the dosage form isfulfilled.

Oromucosal immunotherapy can be regarded as a way of inducing toleranceand inducing mucosal vaccination. The mucosa of the mouth is rich indendritic cells with a strong potential for antigen presentation. Thedendritic cells are believed to process the allergens and then migrateto the local lymph nodes where they present allergen derived peptides toallergen specific T cells. During sublingual immunotherapy thisdendritic cell—T cell interaction is believed to induce T cells withregulatory potential or to increase the ratio of allergen specific Th1cells to allergen specific Th2 cells. A number of immunologicalparameters monitored during the allergy vaccination may be suitablemarkers for effects or efficacy of the treatment, alone or incombination respectively. These include systemic and mucosal antibodyresponses e.g. specific IgA, IgG and IgE antibodies; cytokine levelse.g. INFgamma, IL-2, IL-4, IL-5, IL-10, IL-12 and TNF alpha in blood ormucosal secretions; activation, chemotaxis, proliferation, signaling,cytokine production and other responses of regulatory T-cells, Th1cells, TH2 cells, CD8 cells, other T cell subsets or B-cells or NKcells, and cell surface marker expression such as CD (cluster ofdifferentiation) markers e.g. CD4, CD8, CD23, CD25, CD62L, CLA, beta7,CCR9, CD69, CD45RO, CCR3, CXCR5, effector cell function such as totalhistamine content of basophils; eosinophil, basophil, lymphocyte,monocyte numbers in blood, tissue and secretions; eosinophil, basophil,lymphocyte, monocyte mediator release, cytokine production, activation,chemotaxis, proliferation, signalling and other responses.

In a preferred embodiment a vaccine according to the present inventionhas a profile where one or more of the following immunological changescan be found; an increased allergen specific IgG response, an increasedallergen specific IgA response, reduced allergen specific IgE response,few local side effects; reduced allergen specific effector responses ofeosinophils, basophils, lymphocytes and/or monocytes; induction of Tcells with regulatory potential, increased ratio of allergen specificTh1 cells to allergen specific Th2 cells, induction of other cells withregulatory potential, reduced allergen specific Th2 response.

Allergy is also a known disease in animals especially domestic andcompanion ship animal. It is known in the art that they developallergies toward numerous allergen sources including grass, house dustmites, and parasites. Hematophagous, i.e. bloodsucking insectinfestation is known to lead to a hypersensitive response called fleaallergic dermatitis (FAD). In a preferred embodiment of the currentinvention allergens for animal vaccines include allergens originating ortransferred from parasites like ectoparasites (e.g. fleas, ticks,mosquitoes, flies), parasitic helminth venom (like heart worm e.g.Dirofilaria or onchocerciasis e.g. Onchocerca) and house dust mite. Morepreferred are saliva allergens from fleas like Ctenocephalides e.g. C.canis and C. felis, hard ticks likes Ixodes, Arnblyomma, soft ticks likeOrnithodoros and from midges like Culicoides.

EXAMPLES Abbreviations

API: Active Protein Ingredient

ELISA: Enzyme Linked Immuno Sorbent Assay

DDT: Dithiothreitol

HRP: Horse Radish Peroxidase

LIA: Magic Lite specific IgE assay

LITE-reagent: Luminescence labelled anti-IgE

PMP: Para Magnetic Particles

SDS-PAGE: Sodium dodecyl sulphate poly-acryl amide gel electrophoresis

TMB: Tertamethylbenzidine

Example 1 Allergen Vaccine Containing Phleum pratense Grass PollenExtract and Fish Gelatin

Composition:

TABLE 1 Dosage Dosage Dosage Ingredients Unit form 1 form 2 form 3Function Drug substance: Phleum pratense SQ-U 2500 25000 125000 API mg0.0047 0.047 0.235 extract Other ingredients Purified water mg q.s toq.s to q.s to solvent 250 mg 250 mg 250 mg Gelatine (standard mg 10 1010 Matrix molecular weight fish gelatin, Croda UK) Mannitol mg 7.5 7.57.5 Matrix Sodium mg q.s q.s q.s pH adjust- hydroxide ment to 7.5Grass Extract

Grass pollen extract was prepared according to the method describes inIpsen and Løwensten (1983) Jour. Allergy. Clin. Immunol. 72:2, page150-159. In short grass pollen was extracted in ammonium hydrogencarbonate, for 20 hours at 5° C. Particulate matter was removed bycentrifugation and the supernatant was dialysed against water (3 times),lyophilised and stored cold until reconstitution.

Solid Dosage:

Manufacturing Process:

-   -   1. The mannitol was added to an aliquot of the purified water        (not less than 50% of the total batch requirement) and allowed        to dissolve.    -   2. The gelatine was added to the mannitol solution and the        solution was stirred on a magnetic stirrer until the gelatine        had fully dissolved.    -   3. A second aliquot of the purified water (not more than 35% of        the total batch requirement) was used to reconstitute the        allergen extract in the vials. The reconstituted allergen        extract was added to the mannitol-gelatine solution.    -   4. The pH of the bulk formulation was adjusted to pH 7.5 using        freshly prepared sodium hydroxide solution (3% w/w).    -   5. The additional amount of purified water required to complete        the formulation was calculated and transferred to the bulk mix.    -   6. The solution was dosed into pre-formed blister packs. The        solutions were dosed under ambient temperature conditions.    -   7. After dosing, the filled blister packs were passed through a        liquid nitrogen freeze tunnel. All frozen products were        immediately placed in a frozen storage, prior to freeze-drying.        The units were freeze-dried using standard conditions of shelf        temperature and chamber pressure.    -   8. The freeze dried units were sealed with a lidding foil and        finally packed in a sachet

The solid dosage form had an average weight of 18 mg and averagediameter of 11 mm.

Short Descriptions of Analytical Methods:

Identity (ID), Protein Profile:

The protein profile was determined by SDS-PAGE on a Novex Mini CellXcell II system (Invitrogen) according to manufacturers instructions. Inshort, samples are diluted with sample buffer added reducing agent (0.5M DDT), and subjected to 70° C. for 10 min and let to cool for 5 min.Sample, in-house reference and standard low-range size marker (byBIO-RAD) per well are applied on a NuPAGE 4-12% Bis-Tris gradient gel.Electrophoresis is performed at 200 V for approximately 35 min.Subsequently the gel is stained with silver stained. The protein patternshould be similar to that of the In house reference.

Visual Inspection

All units were subjected to visual inspection e.g. colour, shape,irregularities and defects to ensure acceptable appearance.

Disintegration:

The test was performed as described in the European Pharmacopoeia(3^(rd) edition) or the current USP.

Water Content:

The residual water was determined using a Karl Fischer titrationprinciple. The method gives a quantitative determination of the watercontent in a sample based on the principle that a given amount of I₂leads to transformation of an equivalent amount of H₂O.

In short, 1-3 solid dosage form per vial were tested in triplicates in aKarl Fisher Titrator according to the manufactures instruction togetherwith blind samples (4 per run) and KF standard samples see Example 10.

Total Allergenic Activity:

The test was performed using LIA (described in Eiken et al., Allergy1992, 47:495-497), which is a competitive immunoassay. 100 μl anti humanIgE monoclonal antibody bound to paramagnetic particles (PMP) (ADVIACentaur PMP, ALK-Abelló A/S, Denmark) were washed×3 and 100 μl of a poolof patient sera with specific Phleum pratense IgE antibodies were addedand were incubated on a shaker for 2 hours at 2-8° C., whereby specificIgE binds to the PMP. The PMP were washed to remove IgG antibodies×3with gelatine buffer. Ten solid dosage forms were dissolved in gelatinebuffer and dilutions were prepared of 625 SQ-Units or 1250 SQ-Units pertablet. Samples or references of a known content of biotinylated Phleumpratense API were applied and incubated overnight on a shaking at 2-8°C. The samples and the biotinylated API will compete for the IgE bindingsites, when the concentration of allergen in the sample rise, the amountof bound biotinylated API will drop. After incubation the samples werewashed×3 in gelatine buffer, and LITE-reagent for example, streptavidincoupled acridinium ester chemiluminescent compound (ADVIA Centaur LiteReagens, ALK-Abelló A/S is applied. The samples were incubated for 2hours on a shaker at 2-8° C., washed in gelatine buffer×3 and read in aluminometer. The response is inversely related to the concentration ofthe allergen in the sample.

Major Allergen Content:

The test was performed using ELISA technique according to Obispo et al,Allergy, 1997, 52, pg. 806-813.

The ELISA method measures the concentration of Phleum pratense majorallergen 5 (Phl p 5). Two monoclonal antibodies (ALK-Abelló A/S, DK)reacting with different epitopes on the Phl p 5 molecule were coated tothe microtiterplate the night over at 4° C. After washing (4 times withwashing buffer, 0.1 M PBS, 0.05% Tween-20) and blocking the plate withblocking buffer (2% Caseinbuffer), samples/references, which then bindsto the antibodies, were applied. After washing again (4 times withwashing buffer) biotinylated rabbit polyclonal antibodies (ALK-AbellóA/S, DK) against Phleum pratense antigens were applied to the wells andallowed to react.

After 4 times washing with wash buffer, streptavidin coupled to HRP(horse radish peroxidase) (DAKO, Denmark) was applied to the wells andallowed to react for 1 hour at room temperature (shaking). After washing4 times with washing buffer substrate (TMB, KEM EN TEC) for the HRPenzyme was applied and allowed to react for 20 min, the reaction wasthen stopped with 0.5 N sulphuric acid. The colour developed wasmeasured at 450 nm in a spectrophotometer e.g. Multilabel counter Victor2.

Friability:

The friability of the fast-dispersing dosage forms was measured usingthe following method.

A sealed blister sheet containing 10 blisters, each of the blisterscontaining a solid dosage form, was cut apart into 10 individualblisters and each blister was placed in a piece of equipment suitablefor friability measurements as described in EP 3^(rd) ed. V. 2.9.7 andthe units were rotated 100 turns at 25±1 rpm. The individual blisterswere removed, opened and the solid dosage form was transferred to asuitable container. The solid dosage form was then removed from thecontainer, leaving any loose residues in said container. Animmunochemical allergen specific assay (ELISA) was carried out to detectthe amount of allergen content in the residues (see above).

Stability Results:

TABLE 2 Product Dosage form containing Phleum pratense 2500 SQ-U Tests:Friability % loss of total Major Total content Disinte- Water Visualallergen allergenic Sampling of extract gration content inspec- contentactivity (month) (API) (sec.) (%) tion (%) (%) Storage condition: 25°C./60% RH Start 0.000 8 5.5 Comply 96 101 1 n.m. 5 4.9 Comply 79 91 2n.m. 6 5.4 Comply 96 102 3 0.000 5 5.2 Comply 94 82 6 n.m. n.m. n.m.n.m. n.m. n.m. 9 0.000 5 5.3 Comply 83 105 Storage condition: 40° C./75%RH 1 n.m. 3 4.8 Comply 85 86 2 n.m. 8 5.2 Comply 85 100 3 0.005 6 5.4Comply 94 83 6 n.m. n.m. n.m. n.m. n.m. n.m. 9 0.005 4 5.1 Comply 70 92

TABLE 3 Product Dosage form containing Phleum pratense 25000 SQ-U Tests:Friability % loss of Major Total total content Disinte- Water allergenallergenic Sampling of extract gration content Visual content activity(month) (API) (sec.) ID (%) inspection (%) (%) Storage condition: 25°C./60% RH Start 0.000 8 Comply 6.3 Comply 106  104 1 n.m. n.m. Complyn.m. n.m. n.m. n.m. 2 n.m. n.m. Comply n.m. n.m. n.m. n.m. 3 n.m. n.m.Comply n.m. n.m. n.m. n.m. 6 n.m. 7 Comply n.m. Comply 87 141 9 0.000 4Comply 5.1 Comply 79 105 Storage condition: 40° C./75% RH 1 n.m. n.m.Comply n.m. n.m. n.m. n.m. 2 n.m. n.m. Comply n.m. n.m. n.m. n.m. 30.005 n.m. Comply n.m. n.m. n.m. n.m. 6 n.m. 6 Comply n.m. Comply 84 1359 0.005 4 Comply 5.0 Comply 77 105

TABLE 4 Product Dosage form containing Phleum pratense 125000 SQ-UTests: Friability % loss of Major Total total content Disinte- Waterallergen allergenic Sampling of extract gration content Visual contentactivity (months): (API) (sec.) ID (%) inspection (%) (%) Storagecondition: 25° C./60% RH Start 0.000 5 Comply 4.7 Comply 100  100 1 n.m10  Comply 3.9 Comply 84 93 2 n.m. 5 Comply 4.5 Comply 92 104 3 n.m. 8Comply 4.8 Comply 80 88 6 n.m. 6 Comply n.m. Comply n.m. n.m. 9 0.003 5Comply 4.5 Comply 77 106 Storage condition: 40° C./75% RH 1 n.m. 4Comply 4.1 Comply 90 92 2 n.m. 6 Comply 4.6 Comply 91 109 3 0.001 7Comply 4.5 Comply 83 89 6 n.m. 5 Comply n.m. Comply n.m. n.m. 9 n.m.  5*Comply 4.6 Comply 78 118 *Mean value of only 3 dosage forms

Example 2 Allergen Vaccine Containing Phleum pratense Grass PollenExtract and Starch

Composition:

TABLE 5 Dosage Dosage Dosage Ingredients Unit form 1 form 2 form 3Function Drug substance: Phleum pratense SQ-U 2500 25000 125000 API mg0.0047 0.047 0.235 extract Other ingredients Purified water Mg q.s toq.s to q.s to solvent 250 mg 250 mg 250 mg Pre-gelatinised Mg 8 mg 9 mg11 mg Matrix starch Mannitol Mg 8 mg 9 mg 11 mg Matrix Sodium hydroxideMg q.s q.s q.s pH adjust- ment to 7.5Manufacturing Process:

Same as example 1, pre-gelatinised starch was added instead of gelatine(fish source). The solid dosage form had an average weight of 19 mg andaverage diameter of 11 mm.

Short Descriptions of Analytical Methods:

Same as example 1.

Stability Results:

TABLE 6 Product Dosage form containing Phleum pratense 2500 SQ-U Tests:Friability Total % loss of total Disinte- Water allergenic Samplingcontent of Gration content Visual activity (months): extract (API)(sec.) (%) inspection (%) Storage condition: 25° C./60% RH Start 0.008 83.5 Comply 101  1 n.m. 5 3.0 Comply 80 2 n.m. 6 3.6 Comply 99 3 0.021 53.9 Comply 69 6 n.m. n.m. n.m. n.m. n.m. 9 0.010 5 3.5 Residues 101 Storage condition: 40° C./75% RH 1 n.m. 3 2.9 Comply 74 2 n.m. 8 3.7Comply 98 3 0.022 6 4.3 Comply 73 6 n.m. n.m. n.m. n.m. n.m. 9 0.003 4n.m. Residues 75

TABLE 7 Product Dosage forms containing Phleum pratense 25000 SQ-UTests: Friability % loss of Total total aller- content Disinte- WaterVisual genic Sampling of extract gration content inspec- activity(months): (API) (sec.) ID (%) tion (%) Storage condition: 25° C./60% RHStart 0.022 10 Comply 3.3 Comply 106  1 n.m. n.m. Comply n.m. n.m. n.m.2 n.m. n.m. Comply n.m. n.m. n.m. 3 n.m. n.m. Comply n.m. n.m. n.m. 6n.m. n.m. n.m. n.m. n.m. n.m. 9 0.010 160  n.m. 3.5 Residues 99 Storagecondition: 40° C./75% RH 1 n.m. n.m. Comply n.m. n.m. n.m. 2 n.m. n.m.Comply n.m. n.m. n.m. 3 n.m. n.m. Comply n.m. n.m. n.m. 6 n.m. n.m. n.m.n.m. n.m. n.m. 9 0.050 60 n.m. 3.4 Residues 96

TABLE 8 Product Dosage forms containing Phleum pratense 125000 SQ-UTests: Friability % loss of Total total aller- content Disinte- WaterVisual genic Sampling of extract gration content inspec- activity(months): (API) (sec.) ID (%) tion (%) Storage condition: 25° C./60% RHStart 0.041 11 Comply 2.6 Comply 121 1 n.m 18 Comply 2.4 Residues 102 2n.m. 30 Comply 3.0 Residues 126 3 0.055 28 Comply 3.6 Residues 102 6n.m. n.m. n.m. n.m. n.m. n.m. 9  0.0030 58 n.m. 2.9 Residues  99 Storagecondition: 40° C./75% RH 1 n.m. 58 Comply 2.5 Residues  96 2 n.m. 25Comply 3.1 Residues 121 3 0.033 25 Comply 3.5 Residues  97 6 n.m. n.m.n.m. n.m. n.m. n.m. 9 n.m. 70 n.m. 2.7 Residues 110

Example 3 Allergen Vaccine Containing Grass Extract and Fish Gelatin

TABLE 9 Dosage Dosage Dosage Ingredients Unit form 1 form 2 form 3Function Active substance: Phleum pratense SQ-U 2500 25000 75000 Activesubstance mg 0.0047 0.047 0.141 extract Other ingredients Purified watermg q.s. to q.s. to q.s. to Solvent 250 mg 250 mg 250 mg Gelatine mg 1616 16 Matrix (fish source)* Mannitol mg 14 14 14 Matrix Sodium mg q.s.q.s. q.s. pH adjust- hydroxide ment 7.5Manufacturing Process:

Same as example 1

The solid dosage form had an average weight of 30 mg and averagediameter of 12 mm.

Short Descriptions of Analytical Methods:

Same as example 1, except friability and stability were not measured.

Results of Analysis:

TABLE 10 Test Methods Major allergen Total allergenic Disinte- Watercontent (%) activity (%) gration content Strengths ELISA LIA (sec.) (%)2500 SQ-U/dosage 97 102 1 5.5 form 25000 SQ-U/dosage 100 89 1 5.3 form75000 SQ-U/dosage 94 95 1 5.2 form

The results provide an embodiment of an allergen containing solid dosageform comprising three different doses of grass allergen extract in amatrix forming agent consisting of 6.5% fish gelatin and 5.5% mannitol.The allergen content and allergenic activity in the dosage forms werewithin the acceptable limits (see below) following manufacture. Further,the results shows that all dosage forms have a water content in thepreferred range of 4-7%.

Results

As apparent from Examples 1, 2, and 3 it is possible to manufacturefast-dispersing solid allergen vaccine dosage forms which disintegrateinstantaneously. The loss of total content of extract was found to beacceptable, even though the visual inspection resulted in the detectionof residues in some of the blister packs (predominantly to a higherdegree for starch containing matrixes), the amount of the residue i.e.allergen content loss is within the acceptable limit, i.e. less than 0.5μg extract. Thus, it is possible to manufacture non-compressedfast-dispersing solid dosage forms with low friability containingallergens.

The stability data show that the formulations are stable at roomtemperature and at elevated temperature and humidity for nine months.The allergen content and total allergen activity remained unchanged(within assay variation and according to EP 3^(rd) ed. monograph forAllergen Products; total allergen activity 50-200% of theoretical value,major allergen content 65-135% of theoretical value).

All manufactured batches were subjected to visual inspection and foundwithin the acceptable limit.

Example 4 Allergen Vaccine Compositions

Solid allergen vaccine dosage forms were prepared containing varyingratios of matrix forming agents.

TABLE 11 Solid dosage form containing 75,000 SQ-U Phleum pratense grasspollen extract prepared in different packs % % Pack Load toDisintegration gelatin mannitol type Fracture (Kgf) times (sec.) 4.003.00 5 layer foil 0.158 <2 4.00 3.00 PVC/PVdC 0.199 <2 5.00 3.75 5 layerfoil 0.296 <2 5.00 3.75 PVC/PVdC 0.264 <2 6.00 4.50 5 layer foil 0.342<2 6.00 4.50 PVC/PVdC 0.386 <2 7.00 5.25 5 layer foil 0.491 <2 7.00 5.25PVC/PVdC 0.421 <2

All dosage forms were prepared in blister pack having a unit diameter of12 mm as described previously. All dosage forms disintegrated rapidlyand were robust as assessed by visual appearance, tensile strength andpeak load to fracture.

TABLE 12 Solid dosage forms containing 75000 SQ-Units of Phleum pratensegrass pollen extract in fish gelatine and mannitol. % % Tensile strengthPeak load to Disintegration gelatin mannitol (N/mm⁻²) Fracture (Kgf)times (sec. 5 4 0.239 0.168 <2 6.5 5 0.361 0.265 <2 6.5 5.5 0.425 0.277<2 5 7 0.389 0.239 <2 8 4 0.531 0.308 <2 8 7 0.708 0.465 <2 7 7 0.5430.355 <2 7 5 0.458 0.311 <2 6 4 0.263 0.169 <2 6 7 0.381 0.265 <2

All dosage forms were prepared in blister pack having a unit diameter of12 mm as described previously.

All dosage forms disintegrated rapidly and were robust as assessed byvisual appearance and Peak load to fracture.

Example 5 Uniformity

Dosage forms according to composition and manufacture as described inexample 1 where tested for uniformity of allergen content. The allergencontent was determined as uniformity of potency of grass pollen Phleum p5 by an ELISA assay as described in example 1 for the dosage formcontaining 25000 and 125000 SQ-Units respectively. 10 individual unitsfrom a blister pack were compared as shown in table 13 and 14.

TABLE 13 Uniformity of content of 25000 SQ-Unit dosage form. ID % oftotal (dosage form no.) allergen content 1 97.0 2 98.6 3 97.7 4 95.6 597.2 6 99.3 7 95.7 8 96.9 9 97.4 10 98.3 mean value 97.4

TABLE 14 Uniformity of content of 125,000 SQ-Unit dosage form ID % oftotal (dosage form no.) allergen content 1 101.4 2 102.4 3 102.3 4 101.95 104.4 6 100.5 7 101.1 8 104.0 9 113.4 10 97.1 mean value 102.8

All variations were within acceptable limits and good uniformity ofallergen content was found.

Example 6 Administration of a Phleum pratense Grass Pollen Vaccine toDogs

Dogs were equally distributed in respect to sex within each study groupand were dosed following according to table 15

TABLE 15 Group assignment dose level Number of Number of Number of Group(in SQ-units) dosage forms^(a) dogs recovery dog 1 0 1 8 4 2 25000 1 8 —3 500000 4 8 4 ^(a)Group 1 received placebo dosage forms, group 2received 25000 and group 3 received 125000 SQ-Unit dosage forms preparedaccording to example 1

The dogs were administered doses as indicated in Table 15 sublingually.The dosage form was placed under the tongue and the snout was heldclosed to allow dissolution of the dosage form. The animals were dosedonce per day for a period of 4 consecutive weeks. Blood samples weredrawn for all dogs in every group after the completion of the treatmentperiod. 4 dogs in the placebo and the high dose group respectivelycontinued through a recovery period of 4 weeks where after further bloodsample were drawn.

Method:

Phleum pratense (Phl p) specific IgG in either serum or plasma wasdetermined as follows: ELISA plates (Costar) were coated with 10 μg/mlPhl p extract over night at 4° C. The plates were washed 4 times with 1min soak in between and blocked against unspecific binding with 2%Casein buffer for one hour at room temperature. Individual serum orplasma samples were diluted in polypropylene plates, transferred to theELISA plates and incubated for two hours at room temperature. Afterwashing, HRP marked anti-dog IgG (ICN) was added to the ELISA plates andincubated for one hour at room temperature. After another wash, TMB wasadded to the ELISA plates, covered and incubated for 20 min at roomtemperature. The reaction was stopped with 0.5M sulphuric acid. Theabsorbance (OD) was measured in a spectrophotometer at 450 nm.

The OD values at 1:200 dilution were compared for the dogs in the threegroups: placebo, 25000 SQ/dose and 500000 SQ/dose. Statisticaldifference between the three groups was calculated with the Mann-Whitneyrank sum test which is a nonparametric test that compares two unpairedgroups. Dogs receiving 500000 SQ units had higher mean value than both25000 and placebo indicating a specific antibody response.

Results:

P values from the Mann-Whitney test are depicted in Table 16.

TABLE 16 Groups P value Placebo vs 25000 SQ-U/dose 0.059 Placebo vs500000 SQ-U/dose 0.004 P-level ≦ 0.05 = significant with a 95%certainty.

There is a clear significant difference between the placebo and the500,000 SQ-U/dose group indicating that sublingual treatment with500,000 SQ-U/dose for 4 weeks gives a higher humoral specific IgG level.There is a borderline significant difference between the placebo and25,000 SQ-U/dose group also indicating that treatment with 25,000SQ-U/dose gives a humoral specific IgG level, although weaker thantreatment with 500000 SQ-U/dose.

Example 7 Administration of a Phleum pratense Grass Pollen Vaccine toAllergics

Allergic patients, both female and male aged 18-65 years, with adiagnosis of grass pollen allergy were administered sublingual doses asone, two or three single doses and/or as multiple doses of a grasspollen extract containing solid dosage form, according example 1, in arandomised, double-blind, placebo-controlled designed trial.

Safety/tolerability was assessed with progressing single doses. Singledoses of placebo, 2,500, 25,000, 75,000, 125,000 and 375,000 SQ-U wereadministered stepwise in a dose-escalating fashion using combinations ofplacebo and active tablets (12 mm in diameter and approximately 18 mgdry weight) to give the required dose. Forty-seven patients with allergyto grass pollen were treated. The dosage forms were placed under thetongue and held there for 1 minute before swallowing. Eating anddrinking was prohibited for 5 minutes after application of the soliddosage form. The patients were observed for 2 hours for symptoms. Allside effects were recorded, and after each dose the patients recordedtolerability on a Visual Analogous Scale. The dosage form containing thedoses were found to be well tolerated up to and including 125,000 SQ-Uas adverse events were predominantly mild in severity and limited to‘itching’ phenomena in the mouth and throat. Adverse events were alsoreported in the placebo group. ‘Itching mouth’ was reported morefrequently with increased dose for example, a progression of adverseevents correlated with progressing doses.

Further, safety/tolerability of repeated doses was tested for threeselected doses, 2,500, 25,000 and 75,000 SQ-U, and placebo. Forty-sevenpatients with allergy to grass pollen, distributed in four groups ofcomparable size, received a daily sublingual dosage for a period of 8weeks. Combinations of three tablets containing placebo, 2,500 and/or25,000 SQ-U (12 mm in diameter and approximately 18 mg dry weight) wereapplied to obtain the required doses. Adverse events were recorded andsymptoms were collected in patient diaries. The doses contained in thedosage form were found to be well tolerated in all three activetreatment groups. Adverse events and symptoms were reported morefrequently with increased dose.

Thus, the solid dosage forms tested are feasible for clinical use inboth escalating dose and singular repeated dose therapy

Example 8 Allergen Vaccine Containing Phleum pratense Grass PollenExtract and Fish Gelatine

Solid dosage forms as described according to example 1 table 1 andmanufactured according to example 1 were stored for 12 month at 25°C./60% RH and evaluated by measurement of visual inspection,disintegration, water content, uniformity of mass, identity (proteinprofile), major allergen content and total allergenic activity. All testwere performed as described in example one. An average of adetermination made in duplicate on a pool of 10 tablets is shown unlessotherwise stated.

TABLE 17 Storage conditions: 25° C./60% RH Product: 2500 SQ-U/tabletsTest: Total Major allergenic allergen Water Visual UniformityDisintegration activity. content. content inspection of mass (seconds)(LIA) (ELISA) Friability (%) 0 Comply Comply 6 122% 97% 0.00% 6.1% 12Comply Comply 1 105% 99% N/A 4.8% Test: Total Major ID allergenicallergen Water Visual (SDS- Uniformity Disintegration activity. content.content inspection page) of mass (seconds) (LIA) (ELISA) Friability (%)Product 25000 SQ-U/tablets 0 Comply Comply Comply 5 108% 100% 0.00% 5.7%12 N/A Comply N/A N/A N/A N/A N/A N/A Product 125000 SQ-U/tablets 0comply Comply Comply 7 110%  99% 0.00% 5.4% 12 comply Comply Comply 1119% 104% N/A 4.2%Visual Inspection:

There were no changes in the appearance of the tablets during the study.

Disintegration:

There were no remarkable changes in the disintegration time during thestudy. All test samples disintegrated immediately.

Uniformity of Mass:

There were no remarkable changes in the uniformity of mass during thestudy (assessment of 20 tablets).

Water Content:

There were no remarkable changes in the water content during the study.

Identify SDS-Page:

There were no dramatic changes in the protein profile during the study;the samples were similar to the reference at all test times.

Total Allergenic Activity:

No significant loss of total allergen activity was measured for thetablets (varies from 105%-119% of the theoretical content for thedifferent strengths at 12 month, values which are within the deviationof the test performed).

Major Allergen Content:

No significant loss of allergen content as determined by major allergencontent was measured for the tablets (varies from 99%-104% of thetheoretical content for the different strengths at 12 month storage,values which are within the deviation of the test performed).

Example 9 Allergen Vaccine Containing Phleum pratense Grass PollenExtract and Fish Gelatine

The composition is as described in example 1, table 1

Solid dosage forms manufactured according to example 1 were stored for18 month at 25° C./60% RH and evaluated by visual inspection of thedosage form, i.e. tablets, and by determining the disintegration time,water content, uniformity of mass, identity (protein profile), majorallergen content and total allergenic activity. All tests were performedas described in example 1. An average of a determination made induplicate on a pool of 10 tablets is shown unless otherwise stated.

TABLE 18 Storage conditions: 25° C./60% RH Product 2500 SQ-U/tabletsTest: Total Major allergenic allergen Water Visual UniformityDisintegration activity. content. content inspection of mass (seconds)(LIA) (ELISA) Friability (%)  0 mth Comply Comply 6 122% 97% 0.00% 6.1%18 mth Comply Comply 1 115% 90% N/A 5.0 Product 125000 SQ-U/tabletsTest: Total Major ID allergenic allergen Water Visual (SDS- UniformityDisintegration activity. content. content inspection page) of mass(seconds) (LIA) (ELISA) Friability (%)  0 mth Comply Comply Comply 7110% 99% 0.00% 5.4% 18 mth Comply Comply Comply 1 117% 94% N/A 4.4% Mth= month(s) N/A = not analysed

For solid dosage form containing 25,000 SQ-Unit/solid dosage form onlyanalyses of ID were performed. Similar ID pattern was found after 18months of storage as compared to 0 month (data not shown).

The solid dosage form were found to be stable, i.e. allergen content andtotal allergen activity remained unchanged (within assay variation andaccording to EP 3^(rd) ed. monograph for Allergen Products; totalallergen activity 50-200% of theoretical value major allergen content65-135% of theoretical value), after storage for 18 months at 25° C./60%RH. Further, there were no changes in the appearance of the solid dosageforms for all the tested allergen doses during the study. Further, therewere no changes in the uniformity of mass of the dosage forms testedduring the study. As to the water content, no noticeable changes wereand within the preferred range of 4-7%.

Example 10 Allergen Vaccine Containing Phleum pratense Grass PollenExtract and Fish Gelatin

The composition of the solid dosage forms is shown in table 9 of example1.

The solid dosage forms were prepared according to example 1 andsubmitted to stability testing.

All tests were performed according to example 1.

Stability Results

TABLE 19 Product 2500 SQ-U/dosage form Test: Sampl- Visual Totalallergenic Major allergen Water ing inspec- Disinte- activity. content.content (months): tion gration (LIA) (ELISA) (%) Storage conditions: 25°C./60% RH 0 Comply  1 sec. 129% 82% 5.0% 1 Comply 1 sec  87% 98% 5.3% 3Comply 1 sec 130% 116%  5.2% 6 Comply 1 sec  97% 100%  5.2% Storagecondition: 40° C./75% RH 1 Comply 1 sec  97% 90% 5.2% 3 Comply  1 sec.124% 114%  5.5% 6 Comply 1 sec 101% 93% 5.9%

TABLE 20 Product 25000 SQ-U/dosage form Test: Total aller- Major VisualID genic allergen Water Sampling inspec- (SDS- Disinte- activity.content. content (months): tion page) gration (LIA) (ELISA) (%) Storageconditions: 25° C./60% RH 0 Comply Similar to 1 sec. 101% 93% 4.7%reference 1 n.m Similar to n.m n.m. n.m n.m reference 3 n.m Similar ton.m n.m n.m n.m reference 6 n.m Similar to n.m n.m n.m n.m referenceStorage condition: 40° C./75% RH 1 Comply Similar to n.m n.m n.m n.mreference 3 Comply Similar to n.m n.m n.m n.m reference 6 Comply Similarto n.m n.m n.m n.m reference

TABLE 21 Product 75000 SQ-U/dosage form Test: Total aller- Major VisualID genic allergen Water Sampling Inspec- (SDS- Disinte- activity.content. content (months): tion page) gration (LIA) (ELISA) (%) Storageconditions: 25° C./60% RH 0 Comply Similar to 1 sec. 109%  77% 5.3%reference 1 Comply Similar to 1 sec 75% 92% 5.1% reference 3 ComplySimilar to 1 sec 113%  108%  5.0% reference 6 Comply Similar to 1 sec99% 95% 5.2% reference Storage condition: 40° C./75% RH 1 Comply Similarto 1 sec 79% 92% 5.3% reference 3 Comply Similar to 1 sec. 105%  100% 5.5% reference 6 Comply Similar to 1 sec 99% 95% 5.9% reference n.m. =not measured

The quality of the dosage forms were judged by visual inspection of thedosage form (i.e. tablets) and by determining the disintegration time,water content, uniformity of mass, identity (protein profile), majorallergen content and total allergenic activity (not all data are shown).

The solid dosage form in all doses were found to be stable, i.e.allergen content and total allergen activity remained unchanged (withinassay variation and according to EMEA 3^(rd) ed. monograph for AllergenProducts; total allergen activity 50-200% of theoretical value, majorallergen content 65-135% of theoretical value), after storage for 6months at 25° C./60% RH and 40° C./75% RH. Further, there were nochanges in the appearance of the solid dosage forms for all the testedallergen doses during the study. Further, there were no changes in theuniformity of mass of the dosage forms tested during the study. As tothe water content, no noticeable changes were found during the study. Aslight tendency to an increase in water content during storage at 40°C./75% RH in all doses was seen, though all the test samples were withinthe preferred range of 4-7%.

Example 11 Determination of Water Content and Water Activity in AllergenDosage Forms Containing Phleum pratense Grass Pollen Extract and FishGelatin

Composition

TABLE 22 Name of Formulation P Formulation F ingredient (5.5% fishgelatine) (4% fish gelatine) Function Purified water q.s. to q.s. toSolvent 250.00 mg 250.00 mg Phleum pretense 2500, 25000, 2500, 25000,Active or 75000 SQ-U or 125000 SQ-U substance 0.0047 mg, 0.0047 mg,0.047 mg, 0.047 mg, 0.141 mg 0.235 mg Gelatine 16 mg  10 mg Matrix(fishsource) Mannitol 14 mg 7.5 mg Matrix Sodium q.s. to q.s to pHHydroxide pH 7.5 pH 7.5 modifier

The solid dosage forms were prepared according to example 1.

Water activity of the various solid dosage forms were measured on aRotronic Hygroskop BT-RS1 (Rotronic ag, Switzerland). 10 tablets wereused for each measurement. A stable read out signal indicated thatequilibrium was reached, and the relative humidity was subsequentlyconverted to water activity using the following relationship:

$a_{w} = {\frac{P_{w}}{P_{w}^{*}} = \frac{{RH}_{equ}}{100}}$

where

a_(w) is the water activity of a sample,

P_(w) is the partial water vapor pressure above a sample,

P_(w)* is the water vapor pressure above pure water, and

RH_(equ) (ERH: relative humidity equilibrium) is the relative humidityof a sample.

A 756 Karl Fischer Coulometer with a 774 Oven Sample Processor (Metrohm,Herisau, Switzerland) was used for the determination of the watercontent in the tablets. 1-3 tablets were placed in glass vials andsealed with PTFE-coated caps (Metrohm, Herisau, Switzerland). The samplevials were then placed in the 774 Oven Sample Processor (Metrohm,Herisau, Switzerland), and any moisture present in the samples wasevaporated at a temperature of 130° C. Evaporated moisture wastransferred to the reaction cell containing Hydranal®-Coulomat Ovenreagent (Riedel-de-Haën) by nitrogen gas, and a subsequentquantification of the amount of water released was performed by a KarlFischer titration according to the manufacturer's instructions. Theresults are presented in table 24.

TABLE 23 Water activity and water content of various allergen dosageforms containing Phleum pratense grass pollen extract and fish gelatin.Water activity Water content (%) Sample N Mean SD N Mean SD Placebo, 30.46 0.022 3 5.59 0.060 P Formulation 2500 SQ-U 2 0.45 0.004 3 4.930.071 P Formulation 25000 SQ-U 2 0.41 0.001 3 4.67 0.021 P Formulation75000 SQ-U, 2 0.41 0.005 3 4.66 0.053 F Formulation 2500 SQ-U 1 0.44 — 35.45 0.070 F Formulation 25000 SQ-U 1 0.46 — 3 5.19 0.055 F Formulation125000 SQ-U 1 0.44 — 3 4.80 0.056 SD = standard deviation

All solid dosage forms had a mean water activity between 0.41-0.46,although some had a water content above 5%.

Example 12 Allergen Vaccine Containing Phleum pratense Pollen Extract

Composition

TABLE 24 Composi- Dosage Dosage Dosage tion form 1 form 2 form 3Ingredients % w/w Placebo 25,000 75,00 Gelatin Fish 6.0  14 mg 14 mg 14mg (Norland, Canada) Mannitol 5.08 12.7 mg 12.7 mg 12.7 mg/ API (grass 025,000 SQ-U/ 75,000 SQ-U pollen extract) NaOH qs to Qs to qs to qs to pH7.5 pH 7.5 pH 7.5 pH 7.5 Purified qs to qs to qs to qs to water 250 mg250 mg 250 mg 250 mg Total %/Wet 100% 250 mg 250 mg 250 mg fill weightDried weight 27.7 mg 27.7 mg 27.7 mg

The formulation is manufactured as described in example 1 except thatfish gelatin from Norland, Canada is used. The solid dosage form had anaverage diameter of 13 mm.

TABLE 25 Dose of dosage Friability (% loss of total form content ofextract (API)) 25.000 SQ-units 0.000* 75.000 SQ-units 0.000*

The results provide a further embodiment of an allergen containing soliddosage form comprising two different doses of grass allergen extract inand matrix forming agent consisting of 6.0% fish gelatin and 5.08%mannitol. The dosage forms were tested in a friability test according tothe methods described in example 1. The results show that theformulations are stable in respect of friability as allergen release wasdetected.

The present invention is not to be limited in scope by the specificembodiments described herein. Indeed, various modifications of theinvention in addition to those described herein will become apparent tothose skilled in the art from the foregoing description and theaccompanying figures. Such modifications are intended to fall within thescope of the appended claims.

It is further to be understood that all values are approximate, and areprovided for description.

Patents, patent applications, publications, product descriptions, andprotocols are cited throughout this application, the disclosures ofwhich are incorporated herein by reference in their entireties for allpurposes.

The invention claimed is:
 1. A method for administering an allergen to amammal which comprises: oromucosally administering to a mammal having atleast one clinical allergy symptom an oromucosal dosage form comprisingan effective dose of a major grass pollen allergen, and repeating theadministration of an oromucosal dosage form comprising an effective doseof the major grass pollen allergen at least once daily, wherein therepeat administration does not comprise an up-dosing phase, and whereinthe repeated administration of said major grass pollen allergen reducesthe at least one clinical allergy symptom.
 2. The method of claim 1wherein said effective dose of said major grass pollen allergen has apotency of about 65 to about 17,600 BAU.
 3. The method of claim 1wherein said dosage form comprises a solid, non-compressed,fast-dispersion matrix comprising fish gelatin.
 4. The method of claim3, wherein the fast dispersion matrix comprises mannitol, wherein theratio of said fish gelatin to said mannitol is about 2:20 to about 20:1.5. The method of claim 1, wherein said dosage form comprises an amountof Phleum pratense extract that provides the effective dose.
 6. Themethod of claim 5, wherein the Phleum pratense extract comprises 2.5μg-75 μg of Phl p 5 allergen.
 7. A method for administering an allergento a mammal which comprises: oromucosally administering to a mammalhaving at least one clinical allergy symptom an oromucosal dosage formcontaining an effective dose of an allergen, and repeatedlyadministering an oromucosal dosage form containing an effective dose ofthe allergen to said mammal, wherein the repeat administration does notcomprise an up-dosing phase, and wherein the repeated administration ofsaid allergen reduces the at least one clinical allergy symptom.
 8. Themethod of claim 7 wherein said dose has a potency of about 65 to about17,600 Biological Allergen Units (BAU).
 9. The method of claim 7,wherein said dosage form comprises a solid, non-compressed,fast-dispersion matrix.
 10. The method of claim 9, wherein the fastdispersion matrix comprises fish gelatin and mannitol, wherein the ratioof said fish gelatin to said mannitol is about 2:20 to about 20:1. 11.The method of claim 7, wherein said dosage form is a sublingual dosageform.
 12. The method of claim 7, wherein the effective dose is about 650to about 6,000 BAU.
 13. A method for administering an allergen to amammal which comprises: oromucosally administering to a mammal having atleast one clinical allergy symptom an oromucosal dosage form comprising:about 2.5 μg to about 75 μg of Phl p 5 allergen; about 14 mg fishgelatin; and about 12.7 mg mannitol; repeating the administration of thedosage form at least once daily, wherein the repeat administration doesnot comprise an up-dosing phase, and wherein the repeated administrationof said Phl p 5 allergen induces an adaptive immune response sufficientto reduce the at least one clinical allergy symptom.
 14. The method ofclaim 13, wherein the at least one clinical allergy symptom is reducedin severity or relieved.